CAI-based systems and methods for the localized treatment of ocular and other diseases

ABSTRACT

The subject invention provides CAI compounds and formulations thereof, and methods for their use in the localized treatment of non-life threatening diseases. Formulations of CAI compounds of the subject invention include CAI free base and CAI prodrug microcrystallines, microparticles, emulsions, and the like. The subject invention further provides methods for treating non-life threatening diseases using the CAI compounds of the invention (i.e., novel delivery systems and combination therapies), that are effective and are associated with little or no adverse side effects.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional application No.60/612,683, filed Sep. 24, 2004, and is herein incorporated in itsentirety.

BACKGROUND OF THE INVENTION

5-amino-1,2,3-triazole-4-carboxamide derivatives were originallydiscovered as antiparasitic agents and then subsequently demonstrated tobe antiproliferative agents and potential cancer therapeutics. Thespecific compound,5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide(Formula I illustrated below, shown as the free base form), has beendemonstrated to have antiproliferative and antimetastatic activity thatwas linked to decrease of intracellular calcium by inhibition ofnon-voltage-gated calcium channels. Tyrosine kinase andmetalloproteinase pharmacological mechanistic activities andantiangiogenesis activity relevant to antitumor efficacy have also beendescribed for this compound. The compound of Formula I will be referredto in the following discussion using the acronym CAI(carboxy-amido-triazole) which is generally used to describe thecompound.

Clinical investigations have been conducted with CAI in the treatment oflife-threatening diseases. For example, CAI has been used in thetreatment of a variety of refractory tumors, including prostate cancer,lymphomas, glioblastoma, peritoneal cancer, fallopian tube cancer,epithelial ovarian cancer, advanced renal cell carcinoma, metastaticrenal carcinoma, and non-small cell lung cancer (Bauer, K. S. et al ClinCancer Res. 5:2324-2329, 1999; Kohn, E. C. et al Cancer Res.52:3208-3212, 1992; Kohn, E. C. et al. J Biol Chem.269:21505-21511,1994a; Kohn, E. C. et al. Proc Natl Acad Sci USA.92:1307-1311, 1995; Kohn, E. C. et al. Cancer Res. 56:569-573, 1996;Kohn, E. C. et al. J Clin Oncol. 15:1985-1993, 1997; Kohn, E. C. et al.Clin Cancer Res. 7:1600-1609, 2001.

While these studies using CAI indicate that CAI has intrinsic clinicalefficacy for many cancer types, the systemic oral dosage regimens andformulations used in a clinical setting to date have been associatedwith dose limiting side effects. Moreover, toxic effects (cerebellarataxia, peripheral neuropathy and exacerbation of depression) have beenobserved in clinical studies at doses of CAI required to achievecirculating levels that are within a narrow range or those projectedfrom pharmacological studies to be required for effective inhibition ofpathological neovascularization. Furthermore, a serious side effectassociated with clinical use of CAI by systemic administration has beenthe loss of vision for which two cases have been described (Berlin, J.et al. Clin Cancer Res. 8:86-94, 2002). Therefore, use of CAI ascurrently applied in clinical investigations for cancer is effectivelyprecluded for acute or chronic treatment of non-life threateningconditions, in particular for the treatment of ocular diseases describedherein.

To treat certain ocular diseases, such as age-related maculardegeneration (or AMD), and diabetic retinopathy, or diseases withspecific ocular manifestations, such as von Hippel landau syndrome,therapeutic treatments rely on occlusion of the blood vessels usingeither threshold laser photocoagulation, or subthreshold laser combinedwith a photoactivated dye. However, such treatment requires eitherfull-thickness retinal damage by thermal destruction, or damage tomedium and large choroidal vessels thereby precluding any potentialvisual recovery. Further, the subject is left with a scar and visualscotoma. Moreover, recurrences are common, and visual prognosis is poor.Recent research in the treatment of neovascularization has had the aimof causing more selective closure of the blood vessels, in order topreserve the overlying neurosensory retina. Such strategies have beenused for the treatment of diabetic retinopathy, the leading cause ofblindness among working age adults in Europe and the United States.However, extensive ocular tissue damage can occur after panretinalphotocoagulation, with the visual handicap of more limited peripheralvision and poor night vision. With focal laser treatment,photocoagulation often can further compromise macular blood flow.Alternatively, a variety of molecules are in development or have beenapproved that target angiogenic pathways (e.g. the VEGF pathway). Thus,using antiangiogenic compounds is an alternative to lasering ofpatients.

CAI is an antiangiogenic compound; however, the poor aqueoussolubilities of CAI compounds, as well as reported neurotoxicity forCAI, means that novel methods of administration and targetedadministration of CAI compounds are required for providing safe andeffective doses to treat disease and non-life threatening diseases inparticular.

High local concentrations of CAI may be required to treat acute diseasesymptoms while lower concentrations can be effective as continuationtherapy or prophylactic therapy. Additionally the frequency ofadministration of a formulation of a CAI compound can also be used toensure safe and effective local concentrations to slow vascularoutgrowth. Treatment may be necessary from every week, to every month tofew months, to yearly dosing with appropriate molecules in sustaineddelivery systems.

Posterior segment neovascularization (NV) is the vision-threateningpathology responsible for the two most common causes of acquiredblindness in developed countries: exudative AMD and proliferativediabetic retinopathy (PDR). Currently the only approved treatments forposterior segment NV that occurs during exudative AMD are laserphotocoagulation, photodynamic therapy with VISUDYNE®, and the VEGFbinding oligonucleotide aptamer Macugen®. Laser and photodynamictherapies involve occlusion of affected vasculature which results inlocalized laser-induced damage to the macula. For patients with PDR,surgical interventions with panretinal laser, or vitrectomy and removalof preretinal membranes, or treatment with Macugen® are the onlytreatment options currently available. However, in addition to therecently approved anti-VEGF aptamer oligonucleotide Macugen®; severaldifferent compounds are being evaluated clinically, including, forexample, anecortave acetate (Alcon Research, Ltd.), and Lucentis(ranibizumab, Genentech), Squalamine lactate (Genaera Corporation),LY333531 (Lilly), Cand5 (Acuity Pharmaceuticals), Talaporfin sodium(Light Sciences Corp.), and Fluocinolone (Bausch & Lomb).

Treatments using dosage regimens, routes of administration andformulations of CAI described to date do not have adequate safety fortreating severe proliferative diseases that are non-life threatening.There exists an unfulfilled need for new dosage regimens, routes ofadministration and formulations of CAI that can provide therapeuticeffects on non-life threatening proliferative diseases, as exemplifiedby ocular diseases that are characterized by neovascularization andpathological cellular proliferation and invasion.

BRIEF SUMMARY OF THE INVENTION

The subject invention provides novel sterile aqueous formulations of CAIcompounds, their use in the localized treatment of ocular diseases, aswell as methods for their manufacture. In some embodiments of theinvention, novel and advantageous CAI compounds, their respectiveformulations, and methods for their use are provided. The novel CAIcompounds and aqueous, sterile formulations of the invention areparticularly useful in the localized treatment of non-life threateningdiseases such as proliferative diseases, ocular diseases or conditions,inflammatory conditions, edematous diseases, signaltransduction-mediated diseases, and matrix metalloproteinase-mediateddiseases, and neovascular diseases.

In certain embodiments, CAI compounds of the present invention arecrystalline and small particulate pure free base forms of CAI, CAIprodrugs, and CAI acid addition salt forms. Further embodiments of theinvention provide novel, therapeutically effective formulations ofcrystalline and small particulate pure free base forms of CAI compoundsof the invention.

Specific diseases that can be treated with the CAI compounds of theinvention include, but are not limited to, ocular disease includingneovascular ocular disease, edematous ocular disease, ocular tumors, andintraocular inflammation; dermatological disease including psoriasis,eczema, actinic keratosis, and rosacea; inflammatory disease includingarthritis and arthrosis; neurological diseases includingneurodegenerative disease, pain, and epilepsy; cancers including cancersof the bladder, brain, breast, colon, endometria, kidney, lungs,pancreas, and thyroid; and proliferative vascular disorders includingrestenosis or proliferation associated with angioplasty.

Aqueous, sterile formulations as described herein provide for thelocalized administration of high concentrations of CAI compounds (whichare normaly poorly soluble) for the treatment of such acute therapeuticindications as exemplified by severe neovascular ocular disease stagesof “wet” ARMD or diabetic retinopathy. Similarly, for chronic use, thesubject invention provides sterile, aqueous formulations for thecontrolled release of CAI compounds via oral, systemic or local routesof administration, where such formulations ensure the maintenance oftherapeutic CAI compound drug levels with low variability andreliability for chronic application.

Treating localized proliferative diseases, including those characterizedby neovascular pathology, by the local administration of CAI compoundsof the invention as described herein, provides significant advantages tothe systemic administration of CAI with such systemic administrationhaving been practiced in cancer to date.

Local administration of CAI compounds as provided herein allows for themaintenance of efficacious, non-toxic levels of active CAI free drug atthe local tissue disease site, thus providing a high margin of safetyand enables therapeutically effective treatment of diseases,particularly non-life threatening diseases. The localized treatment ofnon-life threatening diseases using CAI compounds as described hereinprovides significant advantages relative to systemic treatment withforms and formulations of CAI described to date which have unacceptablelow safety thresholds.

Similarly, according to the subject invention, the local administrationof CAI compounds, and formulations thereof, by means of a drug deliverydevice or implant placed in proximity to the local tissue site providesfor the maintenance of efficacious, safe levels of active CAI drugingredient at the local tissue disease site. Thus, the subject inventionprovides a high margin of safety and enables use in treating non-lifethreatening diseases for which CAI dosed systemically, in formsdescribed to date, is practically precluded.

In one embodiment, local administration of CAI compounds as describedherein is suitable for acute disease therapy and induction of remissionof disease signs and symptoms. In another embodiment localadministration of CAI compounds as described herein is suitable forchronic disease therapy and maintenance of remission of disease signsand symptoms.

According to the subject invention, the local ocular administration ofCAI compounds of the invention, and/or formulations thereof, attenuateocular pathological disease processes without unduly compromising normalhealthy ocular function. Thus, local ocular administration of a CAIcompound of the invention, and/or formulations thereof, provides for anefficacious but safe controlled concentration range of CAI directly inthe eye.

Ocular CAI-based therapies, as describe herein, provide significantadvantages for treating neovascular ocular disease relative to currentlaser surgery treatment modalities including panretinalphotocoagulation, which can be accompanied by extensive ocular tissuedamage. In the examples of posterior neovascular ocular diseases, suchas Age Related Macular Degeneration and Diabetic Retinopathy, targetocular pathologies and tissues for treatment are especially localized tothe retinal, choroidal and corneal ocular compartments.

It is contemplated herein that CAI compounds of the invention canreadily penetrate the human scleral tissue after periocularadministration in patients with ocular disease. This particular propertyof CAI compounds and CAI-based formulations of the invention enableeffective minimally invasive delivery of the parent CAI therapeuticentity (or also referred to herein as CAI free base) to the patient'sdiseased ocular tissue compartment(s) in therapeutic concentrationwithout causing ocular toxicity that has been previously observed withthe systemic administration of the biologically active CAI parent drugto cancer patients.

According to the subject invention, tissue targets for localizedtreatment using the CAI compounds include, but are not limited to,dermal, ocular, pulmonary, vascular, joint, nasal, ictic, bone,gastrointestinal, and localized neural tissues e.g. brain and spinalcolumn of the central nervous system. Accordingly, administrations ofCAI compounds of the invention as described herein can be used to treatocular, dermatological, pulmonary, vascular, joint, bone, nasal, ictic,gastrointestinal and neurological diseases. In a preferred embodiment,CAI compounds of the invention, and/or formulations thereof, are usefulfor treating local disease pathologies that are targeted by theestablished pharmacological mechanism(s) of action of CAI to inhibit thepathological increase in intracellular calcium concentration that blockshyperproliferative, inflammatory, neurodegenerative, edematous,macroscopic pathologies associated with localized target diseasesdescribed herein.

In a preferred embodiment, the present invention provides CAI compoundsand CAI compound formulations that provide for controlled release ofpharmacologically active CAI in a targeted tissue and at therapeuticallyeffective concentrations. Such CAI compounds can be locally delivered oradministered to a target tissue in vivo by passive spontaneous,biologically accelerated, or assisted (such as sonophoresis or galvanic,e.g. iontophoresis) mechanisms.

The CAI compounds of the present invention, and formulations thereof,are advantageous because they overcome problems associated withsterility, stability, toxicity, lack of target tissue specificity,safety, efficacy, extent and variability of bioavailability, which existwith the administration of the active form of CAI. The CAI compounds ofthe present invention, and CAI compound formulations, are particularlyadvantageous in minimizing the undesirable side effects associated withcurrent CAI treatment or therapy, such as loss of vision, cerebellarataxia, peripheral neurotropathy, and exacerbation of depression.

As contemplated in the subject invention, where a CAI compound comprisesa CAI prodrug, the CAI prodrug can be converted to a biologically activeCAI compound at a controlled rate via passive (such as by aqueoushydrolysis) or biologically mediated (such as biocatalytic or enzymatic,e.g. by intraocular esterases) mechanisms. An advantage of the in vivoconversion of the CAI-based prodrug to CAI is that the ensuing CAIprovides localized therapeutic effects in target disease tissue withhigh therapeutic margins of safety.

A further embodiment provides the use of CAI compounds in conjunctionwith a drug delivery system in the form of an implant or a device forthe treatment of conditions as set forth herein. Certain embodiments ofthe invention contemplate the use of CAI compounds and formulationsthereof for use comprising a coating for example of a device inconjunction with physical device implants such as stents and bandligatures. Therapeutic uses of such implants include but are not limitedto vascular diseases such as restenosis, and in bone and tissue grafts.

A further embodiment of the subject invention provides for the localadministration of CAI compounds in combination with otherpharmacological therapies. As contemplated in the subject invention,combination therapies of CAI compounds with other medicaments targetingsimilar or distinct disease mechanisms have advantages which includegreater efficacy in therapeutic treatment of a disease and greatermargins of safety relative to respective monotherapies with eitherspecific or separate medicament.

In one embodiment, a CAI compound is used in a method to treatneovascular ocular disease comprising localized (for example, in oculartissue) concurrent administration with one or more other medicamentseach of which may act to block angiogenesis by a pharmacologicalmechanism. Such combinations would act by providing for superiorefficacy from mechanisms of action from additivity, synergy, oramerlioration of side effects. Medicaments that can be concurrentlyadministered with a CAI compound of the invention include, but are notlimited to, vascular endothelial growth factor VEGF blockers (e.g. byVEGF neutralizing binding molecules such as Macugen (Eyetech) andLucentis (ranibizumab, Genentech), Squalamine lactate (GenaeraCorporation); and VEGF tyrosine kinase inhibition) for treatingneovascular ocular disease (AMD and Diabetic Retinopathy) andglucocorticoids (e.g. Triamcinolone) for treating macular edema. Othergrowth factors that can be concurrently administered with the CAIcompounds of the invention (preferably in sterile, aqueous formulations)include, but are not limited to, Pigment Epithelium Derived Factor(PEDF), Erythropoietin (EPO), integrin, endostatin, R-cadherin,angiostatin, and fibroblastic growth factor.

The invention encompasses the use of any of the CAI compounds andformulations thereof in the manufacture of a medicament for the localtreatment of a non-life threatening disease. In a preferred embodiment,the invention encompasses the use of a CAI compound and a formulation,and preferably an aqueous formulation, thereof in the manufacture of amedicament for the local treatment of an ocular disease.

DETAILED DESCRIPTION OF THE INVENTION

The subject invention provides CAI compounds and formulations thereof,and methods for their use in the localized treatment of non-lifethreatening diseases. Formulations of CAI compounds of the subjectinvention can comprise CAI as a free base and a CAI prodrug inmicrocrystalline form, as a microparticle, as an emulsion, and the like.The subject invention further provides methods for treating non-lifethreatening diseases comprising administration to a patient of atherapeutically effective amount of a CAI compound of the invention(i.e., using novel delivery systems and combination therapies), that areeffective and are associated with little or no adverse side effects.

Routes of administration contemplated herein for the delivery of CAIcompounds and/or of formulations thereof by local administrationinclude, but are not limited to, ocular, dermal, nasal, ictic,pulmonary, intravitreal, peribulbar, subtenon, periocular, retrobulbar,subretinal, and posterior juxtascleral subconjunctival routes. Suchroutes of administration may be performed by injection by syringe andusing a needle or cannula or by needle-free systems: topicaladministration via a formulation comprising drops, ointment, and creams,inhalation by means of an inhalation device such as a metered doseinhaler or dry powder inhaler or nebulizer, or in conjunction with drugdelivery systems exemplified by controlled release of a CAI compoundfrom a matrix comprising a contact lenses, (for delivery of drug to atissue of the eye), from a device, from an implant, for release of drugto proximal tissue, and by use of an iontophoretic system e.g., toenhance the rate of penetration of drug through a barrier tissue.

In a preferred embodiment of this invention, a CAI compound, orformulation thereof, is administered locally to ocular tissue via aroute that attenuates ocular pathological disease processes withoutunduly compromising normal or healthy ocular function by acting directlyin the eye (e.g., at the site of diseased tissue) in a controlledtherapeutically effective and localized fashion.

I. Definition of Terms

The term “CAI compound,” as used herein, refers to CAI as a free baseand to CAI analogs, and CAI prodrugs, and to any salts such as acidsalts thereof. The CAI compounds of the invention are particularlyuseful in the treatment of non-life threatening diseases.

The term “therapeutically effective amount,” as used herein, refers tothe amount necessary to elicit the desired biological response such asamelioration or reduction in severity of a symptom by a percentageamount. In accordance with the subject invention, the therapeuticallyeffective amount of a CAI compound is the amount necessary to treatnon-life threatening diseases. In one embodiment, an effective amount ofa CAI compound, or formulation thereof, may ameliorate the severity ofsymptoms and/or complications associated with a non-life threateningdisease. The amelioration in symptom and/or complication severity may bea 5%,10%,15%,20%,25% 30%,35%,40%,45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%,98% or 99% decrease in severity. Preferably, thetherapeutically effective amount of CAI compound locally administered inthe treatment of ocular disease is a dose of 5 mg to 50 mg. Morepreferably 5 mg is dosed intravitreally and 50 mg is dosed periocularly.More preferably, the CAI compound is CAI free base.

The term “patient,” as used herein, describes an organism, includingmammals, to which treatment with the compositions according to thepresent invention is provided. Mammalian species that benefit from thedisclosed compounds and methods of treatment include, but are notlimited to, apes, chimpanzees, orangutans, humans, monkeys; anddomesticated animals (i.e., pets) such as horses, dogs, cats, mice,rats, guinea pigs, and hamsters.

The term “treatment” as used herein covers any treatment of a non-lifethreatening disease in a patient, particularly a human, comprisingadministration of a compound and/or formulation of a compound of thisinvention and methods of the subject invention, and includes:

-   -   I. Preventing the disease from occurring in a patient that may        be predisposed to the disease but has not yet been diagnosed as        having it;    -   II. Inhibiting the disease, e.g., inhibiting occurrence of an        additional disease; or arresting development, inducing        remission, or maintaining remission of the disease;    -   III. Relieving the disease, e.g., causing regression of the        disease or addressing by diminishing or reducing symptoms; or    -   IV. Inhibiting recurrence of a disease.

“Concurrent administration” and “concurrently administering,” as usedherein, includes administering a compound or formulation in atherapeutic method suitable for use with the compounds and methods ofthe invention (for example, administration of a CAI prodrug of ForumulaII or III, below) in the treatment of serious, non-life treateningdiseases.

“Non-life threatening disease(s),” as used herein, includes but is notlimited to non-life threatening proliferative, inflammatory,neovascular, ocular, edematous, signal transduction-mediated diseases,matrix metalloproteinase-mediated diseases, and neurodegenerativediseases. Examples of serious, non-life threatening ocular diseasesinclude, but are not limited to diabetic retinopathy (DR), neovascularage-related macular degeneration (ARMD), diabetic macular edema (DME),cystoid macular edema (CME) and ocular tumors such as retinoblastoma(RB), Retinopathy of Prematurity (ROP), Retinal Vascular Occlusions(RVO), corneal neovascularization, iris neovascularization, neovascularglaucoma, ischemic neural damage, uveitis, glaucoma, and pterygium,neovascular diseases of the retina such as hyperproliferativeretinopathies, vitreoretinopathies, and retinal degeneration associatedwith systemic diseases such as diabetes mellitus, ischemic and hypoxicconditions associated with retinal vein and artery occlusion (e.g., fromsickle cell disease or thrombosis), retinal degeneration resulting fromretinal detachment, and age-related macular degeneration. Non-lifethreatening diseases, as defined herein, include any disease that isdirectly or indirectly mediated by CAI. Non-life threatening diseasesare not diseases which may cause death in a patient. A non-lifethreatening disease may be present in a patient together with alife-treatening disease such as diabetes mellitus.

The present invention may also be used to treat ocular symptomsresulting from diseases or conditions that have both ocular andnon-ocular symptoms. Some examples include AIDS-related disorders suchas cytomegalovirus retinitis and disorders of the vitreous; vonHippel-Lindau Syndrome (a disease that has ocular and nonocularneovascularizations); pregnancy-related disorders such as hypertensivechanges in the retina; and ocular effects of various infectious diseasessuch as tuberculosis, syphilis, lyme disease, parasitic disease,toxocara canis, ophthalmonyiasis, cyst cercosis, and fungal infections.Examples of non-ocular diseases include but are not limited torheumatoid arthritis, psoriasis, contact dermatitis, keratitis,conjunctivitis, scleritis, squamous cell carcinoma, condyloma, eczema,rosacea, vascular proliferation associated with angioplasty, graft vshost disease (organ and tissue transplantation), glioblastoma,peripheral neuropathies, diabetic neuropathy, collagen vasculidities

In addition, the present invention can be used to treat diseases otherthan non-life threatening diseases such as, but not limited to, bladdercancer, breast cancer, brain tumors, colon and rectal cancer,endometrial cancer, kidney cancer, leukemia, lung cancer, melanoma,non-Hodgkin's lymphoma, pancreatic cancer, prostate cancer, skin(non-melanoma) cancer, and thyroid cancer.

The following definitions are applicable to those structures as definedfor Formula II and Formula III, below.

Unless otherwise specified, as used herein, the term “alkyl” refers to astraight or branched chain alkyl moiety. In one embodiment, the alkylmoiety is C₁₋₈alkyl, which refers to an alkyl moiety having from one toeight carbon atoms, including for example, methyl, ethyl, propyl,isopropyl, butyl, tert-butyl, pentyl, hexyl, and octyl.

The term “alkyl” also includes cycloalkyl, including for examplecyclohexyl and the like. The term “alkyl” also includes alkenyl, whichrefers to a straight or branched alkyl moiety having one or more carbondouble bonds, of either E or Z stereochemistry where applicable, andincludes for example, vinyl, 1-propenyl, 1- and 2-butenyl, and2-methyl-2-propenyl.

The term “alkyl” also refers to an alicyclic moiety having from three tosix carbon atoms and having in addition one double bond. Such groups,also known as “cycloalkenyl,” include, for example, cyclopentenyl andcyclohexenyl.

The term “alkyl” also includes alkynyl moieties, including for example,ethynyl, 1-propynyl, 1- and 2-butynyl, 1-methyl-2-butynyl, and the like.

The term “alkoxy” refers to an alkyl-O-group, in which the alkyl groupis as described herein.

The term “halogen” is refers to fluorine, chlorine, bromine and iodine.

The term “aryl” refers to an aromatic carbocyclic ring, e.g., phenyl,substituted phenyl and like groups, as well as rings which are fused,e.g., naphthyl, biphenyl indaryl and the like. Aryl groups may beoptionally substituted with from 1 to 3 groups of alkyl, aryl,heteroaryl, halogen, hydroxy, alkoxy, nitro, cyano, trifluoromethyl,alkylthio, alkylsulfonyl, arylsulfonyl, N(R)₂, S(O)₂N(R)₂, CH₂N(R)₂,CO₂R, and C(O)N(R)₂ wherein R groups are independently hydrogen, alkyl,alkoxyalkyl, hydroxyalkyl, aminoalkyl, arylalkyl, aryl, heterocycloalkylor heteroaryl.

The term “arylalkyl” refers to a moiety in which the “aryl” and “alkyl”groups are as described herein.

The term “heterocycloalkyl” refers to a saturated heterocyclic moietyhaving from two to six carbon atoms and one or more heteroatom from thegroup N, O, and S (or oxidized versions thereof) which may be optionallybenzofused at any available position. This includes, for example,azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, benzodioxolyland the like.

The term “heterocycloalkyl” also refers to an alicyclic moiety havingfrom three to six carbon atoms and one or more heteroatoms from thegroup N, O, and S and having in addition one double bond. Such moietiesmay also be referred to as “heterocycloalkenyl” and includes, forexample, dihydropyranyl, and the like.

In the case that the heterocyclyl moiety contains a nitrogen atom thatmay be substituted, then a substitutent may be chosen from the group ofR.

The term “heteroaryl” refers to monocyclic or bicyclic aromatic ringsystems of five to ten atoms of which at least one atom is selected fromO, N, and S, in which a carbon or nitrogen atom is the point ofattachment, and in which one additional carbon atom is optionallyreplaced with a heteroatom selected from O or S, and in which from 1 to3 additional carbon atoms are replaced by nitrogen heteroatoms. Theheteroaryl group is optionally substituted with up to three groupsselected from alkyl, aryl, heteroaryl, halogen, hydroxy, alkoxy, nitro,cyano, trifluoromethyl, alkylthio, alkylsulfonyl, arylsulfonyl, N(R)₂,S(O₂)N(R)₂, CH₂N(R)₂, CO₂R, and C(O)N(R)₂ wherein R groups areindependently hydrogen, alkyl, alkoxyalkyl, hydroxyalkyl, aminoalkyl,arylalkyl, aryl, heterocycloalkyl or heteroaryl.

This term includes, for example, imidazolyl, tetrazolyl, furanyl,thiophenyl, pyridyl, indolyl, quinolyl, benzothiadiazolyl,benzofurazanyl, benzotriazolyl, and the like.

Certain of the above defined terms may occur more than once in FormulasII and III and upon such occurrence each term shall be definedindependently of the other.

“Optional” or “optionally” means that the subsequently described event,element, or circumstance may or may not occur, and that the descriptionincludes instances where said event occurs and instances in which itdoes not.

The term “substituted” shall be deemed to include multiple degrees ofsubstitution by a named substituent.

Where multiple substituent moieties are disclosed, the substitutedcompound can be independently substituted by one or more of thedisclosed or claimed substituent moieties, singly or plurally.

II. CAI Prodrugs of the Invention

In certain embodiments of the subject invention, CAI prodrugs of FormulaII are provided as follows:

wherein

-   -   R₁ is selected from hydrogen, alkyl, aryl, arylalkyl,        alkylamino, dialkylamino, morpholino, piperazinyl,        alkylpiperazinyl, hydroxy, alkoxy, hydroxyalkyl, and alkoxyalkyl        or is a group of formula        -   wherein X is O, NH or S; Y is selected from hydrogen, alkyl,            alkylamino, dialkylamino, and hydroxyalkyl; R′ is selected            from hydrogen and a naturally occurring α-amino acid side            chain; R″ is selected from hydrogen, alkyl, alkylamino, and            alkylhydroxy.    -   R₂ is selected is selected from hydrogen, alkyl, aryl,        arylalkyl, alkylamino, dialkylamino, morpholino, piperazinyl,        alkylpiperazinyl, hydroxy, alkoxy, hydroxyalkyl, alkoxyalkyl,        alternatively when taken together may form a carbocyclic,        heterocyclic and heteroaryl ring

In certain other embodiments of the subject invention, CAI prodrugs ofFormula III are provided as follows:

wherein

-   -   R₃ is selected from hydrogen, alkyl, aryl, alkylaryl and        heteroaryl or is a group of formula        -   wherein X, Y and Z are independently hydrogen, hydroxy,            alkoxy, amino, alkylamino, dialkylamino, halogen, nitro or            cyano.    -   R₄ is hydrogen, alkyl, aryl, alkylaryl or heteroaryl, or a group        of formula        -   wherein X, Y and Z are independently hydrogen, hydroxy,            alkoxy, amino, alkylamino, dialkylamino, halogen, nitro or            cyano.

Preferred embodiments of CAI prodrugs of Formula II are those thatprovide for a high therapeutic margin of safety via conversion to CAI ata rate that yields localized therapeutic effects in target tissues (suchas ocular tissues, where esterase conversion of an esterified prodrug ofCAI is a preferred embodiment) and compartment of administration. Thepreferred prodrug compounds of the subject invention are particularlyadvantageous because they provide an acceptable safety profileconsistent with acute and/or chronic ocular disease

In a particularly preferred embodiment, CAI prodrugs can readilypenetrate the human scleral tissue and effectively deliver thebiologically active parent CAI therapeutic entity to the ocular diseasetarget tissue compartment(s) in therapeutic concentration withoutcausing ocular toxicity that has been previously described for the CAIparent drug when administered systemically to cancer patients. CAIprodrugs of the present invention may be converted to CAI parent drug,after administration, by both passive (e.g. by pH dependent aqueoushydrolysis) or biologically mediated (e.g. enzymatic) mechanisms.

In certain embodiments, CAI prodrugs of the invention can be convertedin vivo into biologically active CAI with passive hydrolytic mechanisms.For example, a CAI prodrug of Formula II can be converted to CAI in vivoby spontaneous hydrolysis. The rate of spontaneous hydrolysis of CAIprodrugs of Formula II maybe tuned by appropriate choice of the R₁ andR₂ groups. Specifically, to favor the spontaneous hydrolysis ofcompounds of Formula II, one embodiment provides for compounds ofFormula II that contain nucleophilic catalytic groups, such as amino andhydroxyl functions, incorporated into either or both of the R₁ and R₂groups such that internal “anchimeric assistance” of the hydrolyticreaction is catalyzed via formation of a cyclic intermediate with a ringsize ranging from 3-7 atoms.

Alternatively, the conversion of CAI amide prodrugs of Formula II to CAIin vivo may be mediated by endogenous hydrolytic enzymes such asproteases and esterases in the eye. Such hydrolytic enzymes particularlymatrix metalloproteinases are known to be upregulated in targetneovascular disease tissues for the neovascular proliferative diseaseindications of relevance specified herein. CAI amide prodrugs of FormulaII may contain one or more asymmetric carbon atoms and may exist in agiven diastereomeric, racemic, or enantiomeric form.

Inine and amide derivatives of amino functions and are well establishedprodrugs in the medicinal chemistry art. As described above, CAI is aknown organic compound designated as5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamideand having a structure illustrated in Formula I. The amorphous CAI freebase, which is the starting material for crystalline forms, salt forms,and other formulations that are the subject of this invention, isreadily synthesized using previously described methods (see, forexample, U.S. Pat. Nos. 4,590,201 and 5,602,156, the contents of whichare hereby incorporated by reference in their entirety) by a person whois skilled in the art of organic synthesis. CAI amide prodrugs ofFormula II are readily synthesized by persons skilled in the art forexample by reacting CAI with a reactive carboxylic acid derivative (e.g.carboxylic acid chloride or anhydride) in the presence of a base such asan organic base in an organic solvent to promote amide bond formation.Examples of carboxylic acid chlorides and anhydrides include but are notlimited to acetyl chloride, benzoylchloride, propionic anhydride,pivaloyl chloride; examples of organic bases include pyridine,4-dimethylaminopyridine and triethylamine and appropriate organicsolvents include dimethylformamide, tetrahydrofuran and dichloromethane.Compounds of Formula III are readily synthesized by persons skilled inthe art by reacting CAI with the specific ketone or aldehye compound inan organic solvent in the presence of a catalyst or drying agent to formthe CAI ketimine or aldimine prodrugs. Examples of ketones and aldehydesinclude but are not limited to acetaldehyde, proprionaldehyde,benzaldehyde, acetophenone, benzophenone, acetone and ethylmethylketone;organic solvents include dimethylformamide, tetrahydrofuran anddichloromethane; catalysts and drying agents are exemplified bymagnesium sulfate and titanium tetraisopropoxide.

III. CAI Salts of the Invention

Further embodiments of the present invention are novel acid additionsalt forms of CAI free base and CAI prodrugs that are used to prepare apharmaceutical formulation using any of the formulation techniques knownto the skilled artisan or as set forth herein. Particularly preferredare acid addition salt forms that, upon local administration, serve tomaintain a therapeutic, free concentration of CAI that is effective atalleviating disease pathology and have a safety profile consistent withpractical therapeutic application.

Preferred embodiments of the subject invention provide acid additionsalt forms of CAI derived from organic acids that have membranestabilizing or anti-inflammatory effects including, but not limited to,cholic acids, long chain unsaturated fatty acids, and amphilic longchain and sterol derived sulfonic acids. Such salts and complexes may beprepared in a conventional manner and can exhibit superior activity andcontrolled release profile in vivo relative to the simple CAI parentfree base. Such salts refer to non-toxic or biocompatible salts of CAIor CAI prodrugs of the invention that are generally prepared by oneskilled in the art by reacting the CAI in its free base parent form witha suitable organic or inorganic acid.

Representative organic or inorganic acids for the preparation of CAI andCAI prodrug salts of the subject invention include, but are not limitedto, the following: hydrogen borate, hydrogen bromide, hydrogen chloride,nitric acid and hydrogen nitrate, phosphoric acid or trihydrogen ordihydrogen or monohydrogen phosphate, dihydrogenphosphate, sulfuric acidor dihydrogen sulfate, and monohydrogensulfate. Representativehydrophilic organic acid salts include, but are not limited to, thefollowing: citrate, fumarate, gluconate, glutamate, lactate, maleate,mandelate, mesylate, oxalate, succinate tartrate, and valerate.Representative hydrophobic organic acid salts include, but are notlimited to, the following: benzenesulfonate, benzoate, cholate,hydroxynaphthoate, laurate, napsylate, oleate, palmoate, palmitate,salicylate, stearate, tosylate, and taurocholate. Additional salts ofessential amino acids are useful.

Particularly preferred are acid addition salt forms of CAI that, uponlocal administration, serve to maintain a therapeutic concentration offree form CAI that is effective at alleviating disease pathology withouta toxic or tissue-damaging or tissue function-imparing side effectprofile that can prohibit practical therapeutic use. Even more preferredembodiments are acid addition salt forms of CAI derived from orcomprising organic acids that have membrane stabilizing oranti-inflammatory effects such as cholic acids, long chain unsaturatedfatty acids, and amphilic long chain and sterol derived sulfonic acids.

III. CAI Compound Formulations

In a further embodiment, formulations of CAI compounds are provided fortreatment of the non-life threatening diseases, which formulations andmethods of treatment comprise the subject of this invention. Suchformulations serve to overcome bioavailability limitations of CAI as apoorly water-soluble compound and/or provide for the increased, withrespect to an aqueous solution of free base CAI, bioavailability of freeCAI drug to the targeted disease tissue and providing exposure todiseased tissue at a desired rate to achieve and maintain atherapeutically effective amount of CAI in a pharmacologically activeconcentration.

In one embodiment, aqueous CAI compound formulations administered asdescribed herein provide for immediate and complete release of CAI froma formulation. Alternatively, aqueous CAI compound formulations of theinvention, when administered, provide for a slow rate of CAI releasesuch as a controlled release or a zero order release or release of about1% to 5% of the amount of administered drug over a time period of fromabout 1 hour to about 1 month. In related embodiments, formulations ofCAI compounds of the invention include, but are not limited to,microcrystalline forms, micron and submicron solid particulateformulations, emulsion and microemulsion formulations and polymerencapsulation formulations where the drug can be dissolved or suspendedin a matrix comprising a biocompatible and/or biodegradable polymer.

In another embodiment, the CAI compound formulations of the inventionprovide a slow rate of CAI release in the target disease tissue over a1-week to 3-year period after a single administration. Certain CAIcompound formulations of the invention provide pharmacologicallyeffective concentrations of CAI in the target disease tissue over a 15minute to one-week period week after single administration.

The CAI compound formulations described below can be prepared inproportions of ingredients (CAI compound and pharmaceuticly acceptableingredients such as excipitents, surface active agents, solvents, andthe like)of which are determined by the solubility and chemical natureof the CAI compound or formulation, chosen route of administration, andstandard medical practice. For example, microparticle formulations ofCAI can be further prepared in an aqueous formulation, for example inthe presence of a pharmaceutically acceptable surfactant or apharmaceutically acceptable surface active agent.

A. Emulsion Formulations

Emulsion formulations of CAI compounds of the invention include, but arenot limited to, compositions prepared using formulation processescomprising a lipid, surfactant and solvent that are readily performed byone skilled in the art of drug formulation. In a preferred embodimentemulsion formulations of CAI compounds to be administered as describedherein, provide for immediate or short term release of at least 90% ofthe amount of CAI in the formulation and providing pharmacologicallyeffective concentrations of CAI in the target disease tissue over a 15minute to one-week period week after single administration.

In general emulsion formulations of CAI compounds of the invention areprepared using a selection of the following five classes of excipients(Pouton, C. W. Eur. J. Pharm. Sci. 11 Suppl 2: S93-98, 2000): 1) puretriglyceride oils; 2) mixed glycerides; 3) lipophilic surfactants; 4)hydrophilic surfactants; and 5) water-soluble cosolvents. Examples oflipid based drug formulations applicable to CAI, CAI prodrug and saltsthereofinclude emulsions prepared for the lipophilic immunosuppressivedrug cyclosporine (Fahr, A. et al. Clin. Pharmacokinet. 24, 472-495,1993; Fricker, G. et al. Br. J. Pharmacol. 117,217-223, 1996), andhalofantrine a lipophilic small molecule antimalaria drug (Porter, C.J., et al. J. Pharm. Sci. 85, 357-361, 1996).

In certain embodiments, emulsion formulations of the inventioncomprising liquid droplets of formulation comprising CAI or a CAIcompound, which droplets are surface stabilized dispersions of dropletsof sizes less than 10 micrometers in aqueous medium. Alternatively, thesubject invention's emulsion formulations can be locally delivered to atarget tissue as an oil-in-water microemulsion of droplet sizes lessthan 100 nanometers. These microemulsions are composed of CAI, CAIprodrugs and salts thereof dissolved in a mixture of lipophilic vehicleand admixed with surfactants and cosurfactants.

In other embodiments, both these end results (lipophilic carrier andadmixture with surfactants and cosurfactants) are achieved from apreconcentrated or a concentrated solution of a CAI compound in avehicle that forms oil-in-water emulsion of droplet sizes less than 10micrometers spontaneously upon mixing with bodily fluids (i.e., lacrimalfluid, vitreal fluid, scleral fluid, blood, cerebral spinal fluid,vaginal fluid, mucous). Alternatively, emulsion formulations of theinvention can be achieved from a preconcentrated or a concentratedsolution of a CAI compound in a vehicle that forms oil-in-watermicroemulsion of droplet sizes less than 100 nanometers spontaneouslyupon mixing with bodily fluids. In this way in-situ generation of thesmall size droplets is facilitated wherein application of energy is notrequired.

Emulsion formulations of CAI compounds are generally selected andprepared as described below. First, suitable pharmaceutically acceptableoil or triglyceride vehicles preferably a readily availablepharmaceutically acceptable oil or pharmaceutically acceptabletriglyceride vehicle (such as pure triglyceride oils) is selected from acommercial source e.g. soybean oil (Fluka, Buchs, Switzerland), sesameoil (Fluka, Buchs, Switzerland), Miglyol® (810, 812, 818, Hulls,Puteaux, France) and other candidates selection of which is preferablybased on respectively highest solubility and longest term of stabilityof the pure CAI, CAI salts or prodrugs as active ingredient in the oil,solubility and stability determined in preformulation evalution bydissolving CAI in the vehicle, ±surfactant, ±water (optionallycontaining a buffer salt) to determine saturation concentrations of CAIas a function of temperature and then determining the stability of CAIas a function of time (1 day, 2 days, 4 days, 1 week 2 weeks, 4 weeks, 1month, 2 months, and monthly thereafter, each at constant temperaturesof about 20° C., 30° C., and 40° C. Formulations containing hydrophobicsurfactants and hydrophilic surfactants are then prepared with an oilvehicle composition range of 25-45%, hydrophobic surfactant(s) range of25-45% and hydrophilic co-surfactant(s) in a range of 10-20%.

Contemplated hydrophobic surfactants for use in the preparation ofemulsion formulations of the invention include, but are not limited to,glyceryl monostearate and Pluronic® (e.g., Pluronic®F68, Pluronic®F127,Sigma, St. Louis Mo.). Contemplated hydrophilic co-surfactants includebut are not limited to, Tween (e.g., Tween 20, Tween 40, Tween 80,Sigma, St. Louis Mo.) and Labrasol® (Gattefosse, Saint Priest, France).Preferred emulsion formulations of the invention exhibit small submicrondroplet size, high CAI active ingredient stability, and high CAI activeingredient load.

Alternatively, in accordance with the subject invention, a CAI compoundcan be formulated as an aqueous suspension of surface stabilizedparticles. The particle size distribution of such suspensionformulations is carefully maintained to less than 10 micrometers so thatlocal irritation to the target tissue (for example, tissue of the eye)is avoided. For example, one embodiment of the invention provides apreparation of concentrated solution of a CAI compound. Specifically,the CAI compound is dissolved or suspended directly in vehicles thatform a suspension of surface stabilized CAI compound with particles ofsizes less than 10 micrometers. The vehicles provide therapeuticallyeffective concentrations of pharmacologically active CAI at the targetdisease site upon local administration to the disease target tissue ortissue compartment by administration methods described herein. Surfacestabilization of the particles in these embodiments is provided by apharmaceutically acceptable surface active agent, for example a surfaceactive agent selected from the group consisting of lecithin, charged oruncharged phospholipids, polymeric surfactants, non-polymericsurfactants, charged surfactants (e.g., anionic surfactants or cationicsurfactants), uncharged surfactants (e.g., polyether surfactants such aspolyethyleneglycol-containing surfactants andpolyethyleneglycol-co-polypropyleneglycol-containing surfactants), oneor more of bile acids and their salts, or combinations thereof.

Alternatively, the CAI compounds of the invention can be prepared as amolecular complex with beta-cyclodextrin or its derivatives.Furthermore, an alternative formulation consists of molecular complex ofa CAI compound with one or more pharmaceutically acceptable lipidsselected from a group of lecithin, charged or uncharged phospholipids,polymeric surfactants, non-polymeric surfactants, charged surfactants,uncharged surfactants, one or more of bile acids and their salts,triglycerides, or other lipophilic solvents or combinations thereof.

B. Microparticle Formulations

“Microparticlulate forms of a CAI compound” refers to solid particulateforms of CAI free base, CAI prodrugs and salts thereof with particlesizes of 10 micrometers or less. Further, “microparticlulate forms of aCAI compound” refers to respective amorphous or crystalline forms CAI,CAI prodrugs and salts thereof as well as mixtures of CAI, CAI prodrugsand salts thereof with other additives (e.g. polymers, surfactants) thatyield particulate forms of 10 micrometers or less.

Microparticle formulations of CAI compounds of the invention include,but are not limited to, compositions prepared using controlledprecipitation formulation processes readily selected and implemented byone skilled in the art of drug formulation. for example by dissolving aCAI compound in a solvent such as acetone or ethanol or tetrahydrofuranor DMSO to form a solution and then adding said solution to anon-solvent such as water in which the solvent has solubility, or addingsuch non-solvent slowly with stirring to the solution, each methodoptionally in the presence of a pharmaceutically acceptable surfaceactive agent or combination of agents in either the solvent ornon-solvent or in both, followed by removal of the solvent, such as byevaporation or diafiltration to provide a suspension of microparticlesin an aqueous medium. The particles can be isolated by lyophilization ofthe suspension, for example in vials optionally in the presence of anoxygen-free gas such as nitrogen or argon after removal of the water,optionally after addition of a carbohydrate such as lactose or mannitolwhich can form a solid matrix in which the solid particles aredispersed. The particles can be resuspended into an aqueous medium uponaddition of such aqueous medium. In a preferred embodimentmicroparticulate formulations of a CAI compound to be administered asdescribed herein, provide for immediate release of CAI providingpharmacologically effective concentrations of CAI in the target diseasetissue over a one-week to three-year period period after a singleadministration.

In one embodiment, microparticles of CAI of the invention includepolymeric microspheres that encapsulate CAI, CAI prodrugs and saltsthereof of the invention. Microparticle formulations of CAI include, butare not limited to, compositions prepared using controlled precipitationformulation processes readily selected and implemented by one skilled inthe art of drug formulation. A controlled microparticle precipitationprocess is exemplified as follows.

CAI free base or a respective CAI acid addition salt form or prodrug isdissolved in a suitable organic solvent (e.g. ethanol,N-methylpyrrolidine) and then mixed with stabilizers, as exemplified bySPG (0.218 sucrose; buffer salts comprising 0.0038 M monobasic, 0.0072 Mdibasic potassium phosphate; 0.0049 M potassium glutamate). Polymer(e.g. polyvinylpyrrolidone) and sugar (e.g. lactose) matrix-formingcomponents are optionally added. Micron and submicron microparticles ofCAI formulation are then prepared by controlled precipitation at anoptimized temperature under low frequency sonication followed.Sonication of the contents of the vessel containing the components ofsuch formulation can also be accomplished using ultrasound frequencies,for example by placing the vessel containing the formulation in anultrasound bath and applying ultrasound energy. The CAI microparticleformulation(s) are collected, for example by centrifugation such as at10,000 g and characterized by digital optical light microscopy and laserlight scattering and Scanning Electron Microscopy (SEM) for particlesizing. Contents of the formulation are analyzed, for example by usingHPLC for content and stability of such content, and by using USPdissolution techniques to measure the release rate of the CAI drug invitro.

In yet another embodiment, the CAI microparticle formulation is preparedusing methods that are standard in the art, such as by entrapment orencapsulation of CAI free base, CAI salt form or prodrug drug moleculeswithin a matrix of polymeric microspheres of sizes less than 50micrometers, wherein the polymeric matrix material is selected from thegroup of albumin, polylactic-coglycolic acid, polymethacrylic acids andsalts, polyethyleneglycol, natural polymeric materials, syntheticpolymeric materials, charged polymeric materials, and unchargedpolymeric materials, and combinations thereof.

Preferred polymer encapsulation systems include polylactic-coglycolicacid (PLGA) microsphere formulations as exemplified by the formulationof octreotide in Sandostatin-LAR® (See Sandostatin LAR® prescribinginformation (www.sandostatin.com)) which is administered as aonce-a-month depot injectable therapeutic.

C. Liposomal Formulations:

In addition, the compounds of the present invention can also beadministered in the form of liposomes or the like. Disintegratorsinclude, without limitation, delivery systems such as small unilamellarvesicles, large unilamellar vesicles and multilamellar vesicles.Liposomes can be formed from a variety of phospholipids, such aslecithin and phosphatidylcholines, optionally with added cholesteroland/or stearylamine.

D. Pharmaceutical Formulations in General

CAI compounds of the invention may be provided in formulations thatcontain pharmaceutically acceptable auxiliary substances as required toapproximate physiological conditions, such as pH adjusting for exampleto adjust pH of an aqueous medium, for example to about pH 7.4 andbuffering agents to maintain such pH, tonicity adjusting agents whichcan be useful, for example, to adjust tonicity of a formulation tophysiological ionic strength, wetting agents and the like, for example,sodium acetate, sodium lactate, sodium chloride, potassium chloride,calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.

The concentration of the CAI compounds of the invention in theirrespective pharmaceutical formulations can vary widely (% w/v), i.e.,from less than about 0.1%, usually at or at least about 2% to as much as20% to 50% or more by weight, and will be selected primarily by desireddosage levels, fluid volumes, viscosities, etc., in accordance with theparticular mode and route of administration selected and target tissue.Preferably, the concentration of CAI compound in the aqueous,therapeutic formulations of the invention range from about 0.01 mg/mL toabout 100 mg/mL. More preferably, the concentration of CAI compound inthe aqueous, therapeutic formulations of the invention for the localizedtreatment of ocular diseases range from about 1 mg/mL to 60 mg/mL offormulation.

The pharmaceutical formulations of the invention may includedelivery-enhancing agents that can be used alone, in combination witheach other, or in combination with another delivery-enhancing agent. Theterm “delivery enhancing agents” includes agents that facilitate thetransfer CAI to the target cell. Examples of such delivery enhancingagents include, but are not limited to, fatty acid esters, surfactants,detergents, alcohols, glycols, surfactants, bile salts, heparinantagonists, cyclooxygenase inhibitors, hypertonic salt solutions,acetates.

As contemplated herein, alcohols that can act as delivery enhancingagents include, without limitation, the aliphatic alcohols such asethanol, N-propanol, isopropanol, butyl alcohol, acetyl alcohol. Glycolsthat can act as delivery enhancing agents in accordance with the subjectinvention include, but are not limited to, glycerine, propyleneglycol,polyethyleneglycol and other low molecular weight glycols such asglycerol and thioglycerol. Acetates such as acetic acid, gluconic acid,and sodium acetate are further examples of delivery-enhancing agents.Hypertonic salt solutions like 1M NaCl are also examples ofdelivery-enhancing agents.

Examples of pharmaceutically acceptable surfactants that can be providedin pharmaceutical formulations of the invention include, withoutlimitation, sodium dodecyl sulfate (SDS) and lysolecithin, polysorbate80, nonylphenoxypolyoxyethylene, lysophosphatidylcholine,polyethylen-glycol 400, polysorbate 80, polyoxyethylene ethers,polyglycol ether surfactants and DMSO. Bile salts such as taurocholate,sodium tauro-deoxycholate, deoxycholate, chenodesoxycholate, glycocholicacid, glycocheno-deoxycholic acid and other astringents like silvernitrate may be used. Heparin-antagonists like quaternary amines such asprotamine sulfate may also be used. Cyclooxygenase inhibitors such assodium salicylate, salicylic acid, and non-steroidal antiinflammatorydrug (NSAIDS) like indomethacin, naproxen, diclofenac may be used.

Examples of pharmaceutically acceptable detergents that can be providedin pharmaceutical formulations of the invention can be selected fromanionic, cationic, zwitterionic, and nonionic detergents. Exemplarypharmaceutically acceptable detergents include but are not limited totaurocholate, deoxycholate, taurodeoxycholate, cetylpyridium,benalkonium chloride, ZWITTERGENT-3-14 detergent, CHAPS(3-{(3-Cholamidopropyl) dimethylammoniol}-1-propanesulfonate hydrate,Aldrich), Big CHAP, Deoxy Big CHAP, TRITON X-100 detergent, C12E8,Octyl-B-D-Glucopyranoside, PLURONIC-F68 detergent, TWEEN-20 detergent,and TWEEN-80 detergent.

The concentration of a delivery-enhancing agent in a pharmaceuticalformulation of the invention will depend on a number of factors known toone of ordinary skill in the art. Such factors include the particulardelivery-enhancing agent being used, the buffer, pH, target tissue ororgan and mode of administration. In certain embodiments of theinvention, the concentration of the delivery-enhancing agent will be inthe range of 1% to 50% (v/v), preferably 10% to 40% (v/v) and mostpreferably 15% to 30% (v/v). Preferably, the detergent concentration inthe final formulation administered to the patient is about 0.5 to about2.times. the critical micellization concentration (CMC).

Excipients can be included in the formulations of the invention.Examples include cosolvents, surfactants, oils, humectants, emollients,preservatives, stabilizers and antioxidants. A pharmacologicallyacceptable buffer may be used, e.g., tris or phosphate buffers.Effective amounts of diluents, additives and excipients are thoseamounts which are effective to obtain a pharmaceutically acceptableformulation in terms of solubility, stability, and biological activity.

Formulations of the invention may also include, depending on theformulation and mode of delivery desired, pharmaceutically-acceptable,non-toxic carriers or diluents, which include vehicles commonly used toform pharmaceutical compositions for animal or human administration. Thediluent is selected so as not to unduly affect the biological activityof the combination. Examples of such diluents which are especiallyuseful for injectable formulations are water, the various commerciallyavailable pharmaceutically-acceptable saline, organic or inorganic saltsolutions, Ringer's solution, dextrose solution, and Hank's solution. Inaddition, the pharmaceutical composition or formulation may includeadditives such as other carriers; adjuvants; or nontoxic,nontherapeutic, nonimmunogenic stabilizers and the like.

Carrier employed may be, for example, either a solid or liquid. Examplesof solid carriers include lactose, terra alba, sucrose, talc, gelatin,agar, pectin, acacia, magnesium stearate, stearic acid and the like.Examples of liquid carriers include syrup, peanut oil, olive oil, waterand the like. Similarly, a carrier for oral use may include time delaymaterial well known in the art, such as glyceryl monostearate orglyceryl distearate alone or with a wax.

In certain embodiments, pharmaceutical formulations comprise a CAIcompound (preferably, CAI itself) and a carrier such as inclusioncompound host materials. The “inclusion compound host materials” asdescribed herein, interact with the CAI compound to increase aqueoussolubility, increase chemical stability, and/or enhance drug (such asCAI compound) delivery to and through biological membranes. It isbelieved that by providing a carrier such as inclusion compound hostmaterials, a stabilized CAI compound molecule can be safely delivered toa patient at a dosage that will not induce toxicity. In addition, suchcarrier materials can include coating materials (i.e., enteric-coatings)that allow dissolution of the coating in an alkaline environment such asin the intestines.

Inclusion compound host materials that can be used in accordance withthe subject invention include those disclosed in U.S. Patent ApplicationNo. 20040033985, incorporated herein by reference in its entirety.Contemplated inclusion compound host materials include proteins (such asalbumin), crown ethers, polyoxyalkylenes, polysiloxanes, zeolites,cholestyramine, colestipol, colesevelam, colestimide, sevelamer,cellulose derivatives, dextran derivatives, starch, starch derivatives,and pharmaceutically acceptable salts thereof. Contemplated cellulosederivatives and dextran derivatives include DEAE-cellulose,guanidinoethylcellulose, or DEAE-Sephadex. Favorable starches or starchderivatives to be included in the compositions of the invention includecyclodextrin, retrograded starch, degraded starch, a combination ofretrograded and degraded starch, hydrophobic starch, amylase,starch-diethylaminoethylether, and starch-2-hydroxyethylether.

According to the subject invention, aqeous CAI compound formulationscomprise CAI and inclusion compound host materials selected from, butnot limited to, cyclodextrin and/or its derivatives (i.e., methylβ-cyclodextrin (M-β-CD), hydropropyl β-cyclodextrin (HP-β-CD),hydroethyl β-cyclodextrin (HE-β-CD), polycyclodextrin, ethylβ-cyclodextrin (E-β-CD) and branched cyclodextrin. As one skilled in theart will appreciate, any cyclodextrin or mixture of cyclodextrins,cyclodextrin polymers, or modified cyclodextrins can be utilizedpursuant to the present invention. Cyclodextrins are available fromWacker Biochem Inc., Adrian, Mich. or Cerestar USA, Hammond, Ind., aswell as other vendors. Formation of inclusion complexes usingcyclodextrin or its derivatives protects the constituent (i.e., CAIcompound) from loss of evaporation, from attack by oxygen, acids,visible and ultraviolet light and from intra- and intermolecularreactions.

The general chemical formula of cyclodextrin is (C₆O₅H₉)_(n). Thecontent of inclusion compound host materials in compositions of thesubject invention can range from about 1 to 80 wt %. Preferably, thecontent of inclusion compound host materials in compositions of theinvention range from about 1 to 60 wt %. The actual amount of theinclusion compound host materials used will depend largely upon theactual content of CAI compound and therapeutic agents, if any, used inpreparing compositions of the invention.

Preferably, aqeous pharmaceutical formulations of CAI compounds can beprepared as molecular complexes with beta-cyclodextrin or itsderivatives. Furthermore, an alternative pharmaceutical formulationconsists of molecular complex of CAI compound(s) with one or more lipidsselected from a group of lecithin, charged or uncharged phospholipids,polymeric surfactants, non-polymeric surfactants, charged surfactants,uncharged surfactants, one or more of bile acids and their salts,triglycerides, or other lipophilic solvents or combinations thereof.

The CAI compound and aqeous formulations thereof of the subjectinvention can be formulated according to known methods for preparingpharmaceutically useful compositions. Formulations are described in anumber of sources, which are well known and readily available to thoseskilled in the art. For example, Remington's Pharmaceutical Science(Martin EW [1995] Easton Pennsylvania, Mack Publishing Company, 19^(th)ed.) describes formulations that can be used in connection with thesubject invention. Formulations suitable for parenteral administrationinclude, for example, aqueous sterile injection solutions, which maycontain antioxidants, buffers, bacteriostats, and solutes, which renderthe formulation isotonic with the blood of the intended recipient; andaqueous and nonaqueous sterile suspensions, which may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in a freeze dried (lyophilized) conditionrequiring only the condition of the sterile liquid carrier, for example,water for injections, prior to use. Extemporaneous injection solutionsand suspensions may be prepared from sterile powder, granules, tablets,etc. It should be understood that in addition to the ingredientsparticularly mentioned above, the formulations of the subject inventioncan include other agents conventional in the art having regard to thetype of formulation in question.

Certain embodiments of the invention may be formulated for localinstillation in or around tissue or organs, where for specific targetedlocal therapy for non-ocular body sites, parenteral therapy is used toachieve safe and effective local drug concentrations into a definedtarget lesion or disease area, or tissue(s), or organ(s), or bodycompartment(s), wherein routes of administration of formulations of CAIcompounds can include transcutaneous, subcutaneous, intradermal,intrathecal, intracerebellar, intramuscular, intra-articular orintravenous, when such direct application is practical and clinicallyindicated.

Alternatively, certain embodiments of the invention may be formulatedfor systemic administration to a patient diagnosed with a non-lifethreatening disease.

The CAI compounds of the invention can be administered in topicalformulations or polymer matrices, hydrogel matrices, polymer implants,or encapsulated formulations to allow slow or sustained release of thecompositions. A particularly preferred formulation is a suspension orsolution of the delivery system in a topical ocular formulation, such aseye drops.

Topical applications may be formulated in carriers such as hydrophobicor hydrophilic bases to form ointments, creams, lotions, in aqueous,oleaginous or alcoholic liquids to form paints or in dry diluents toform powders. Such topical formulations can be used to treat oculardiseases as well as inflammatory diseases such as rheumatoid arthritis,psoriasis, contact dermatitis, delayed hypersensitivity reactions andthe like.

Certain embodiments of the invention are designed to locally deliverbioactive CAI into target eye tissue from implants to be injected in oraround the eye, for sustained delivery of bioactive CAI to the eye.Alternatively, delivery vehicles are designed in the form of ointmentsor gels or viscous suspensions or viscous emulsions that releases thedrug upon administration into or around eye for prolonged periods toelicit the desired pharmacological action. The dosage form can also be afree flowing sterile suspension for topical administration or freeflowing sterile suspensions for injection into or around eye.

When the CAI compound is formulated as a solution or suspension, thedelivery system is in a pharmaceutically acceptable carrier, preferablya pharmaceutically acceptable aqueous carrier, suitable for local tissuecompatibility. A variety of aqueous carriers may be used, e.g., water,buffered water, 0.8% saline, 0.3% glycine, hyaluronic acid and the like.Such delivery systems may be sterilized by conventional, well knownsterilization techniques e.g., by heat or steam sterilization offormulations in sealed vials at about 130° C. for at least 15 minutes,or liquid formulations may be sterile filtered, e.g., by filtrationthrough a 0.2 micron filter in a sterile pharmaceutic environment.

IV. Routes of Administration

The CAI compounds of the invention, and formulations thereof, can beused in a variety of routes of administration, including, for example,orally-administrable forms such as tablets targeting a specific localtissue e.g. gastrointestinal tissues, capsules or the like, or viainjectable (e.g., parenteral, ocular, intravenous, subcutaneous,intramuscular), topical (e.g.,dermal, ocular, transdermal, nasal),buccal, pulmonary (e.g., inhalation), suppository, or other route. Inone embodiment, they can be in unit dosage form, namely, in physicallydiscrete units suitable as unitary dosages for human consumption, eachunit containing a predetermined quantity of active ingredient calculatedto produce the desired therapeutic effect in association with one ormore pharmaceutically acceptable other ingredients, i.e., diluent orcarrier.

The preferred routes of localized administration for the treatment ofocular diseases using the CAI compound or formulations thereof, of theinvention include ophthalmic artery administration, subretinalinjection, intravitreal injection, and periocular injection orjuxtascleral administration. In the case of angiogenic diseases such asage related macular degeneration and diabetic retinopathy, theformulations of the present invention are administered over a course oftreatment ranging from weeks to years. The preferred routes ofadministration for the treatment of such diseases include ophthalmicartery administration, subretinal injection, intravitreal injection, andperiocular injection or juxtascleral administration. Sustained releaseformulations such as implants would also be appropriate for thetreatment of such long term disease indications. These formulations mayalso be administered in combination with other anti-angiogenic agents.

In one embodiment of the invention, a CAI compound can be injectedintraocularly using intravitreal (into the vitreous), subconjuctival(into the subconjuctival), subretinal (under the retina), or retrobulbar(behind the eyeball) injection. For subconjuctival injection, a CAI dosein the range of about 0.1 ng/ml to about 10 mg/ml may be used. Forintravitreal injection, a CAI dose in the range of about 0.1 ng/0.1 mlto about 10 mg/0.1 ml may be used. For retrobulbar injection, a CAI dosein the range of about 1 ng/ml to about 10 mg/ml may be used. Forsubretinal injection, a CAI dose in the range of about 0.1 ng/0.1 ml toabout 10 mg/0.1 ml may be used.

A. Time Released Delivery Route

Slow or extended-release delivery systems, such as delivery systemscomprising a biopolymer (biological-based systems), liposomes, colloids,resins, and other pharmaceutical acceptable polymeric delivery systemsor compartmentalized reservoirs, can be utilized with the compositionsdescribed herein to provide a continuously releasing or long termreleasing source of therapeutic compound. Such slow release systems areapplicable to formulations for delivery via topical, intraocular, oral,and parenteral routes. Delivery to areas within the eye, in situ can beaccomplished by injection such as by using a needle and syringe, by useof a cannula or other invasive device designed to introduce preciselymetered amounts of a desired formulation to a particular compartment ortissue within the eye (e.g. anterior or posterior chamber, uvea orretina). A solid, semisolid, or liquid implant can be deliveredsubretinally using the instrumentation and methods described in U.S.Pat. Nos. 5,817,075 and 5,868,728, the contents of which are herebyincorporated by reference in their entirety.

In a time-release formulation, the microsphere, capsule, liposome, etc.may contain a concentration of CAI that could be toxic if it wereadministered as a bolus dose. The time-release administration, however,is formulated so that the concentration released over any period of timedoes not exceed a toxic amount. This is accomplished, for example,through various formulations of the vehicle (coated or uncoatedmicrosphere, coated or uncoated capsule, lipid or polymer components,unilamellar or multilamellar structure, and combinations of the above,etc.). Other variables may include the patient'spharmacokinetic-pharmacodynamic parameters (e.g., body mass, gender,plasma clearance rate, hepatic function, genetic makeup, etc.).Depending upon the amount of CAI provided in the formulation and therelease rate of the CAI, a patient could be dosed with CAI over a periodof years from a single implant or injection comprising a CAI compound.As illustrative but non-limiting examples, a capsule or a device, e.g.,for implantation use, can be loaded with a CAI-based formulationcomprised of a concentration within the range of 1 to 200 mg of activeCAI ingredient (e.g., 1 mg, 2 mg, 5 mg, 10 mg, 40 mg, 80 mg, 100 mg, 121mg, 155 mg, 200 mg); if the capsule is formulated to release a few(e.g., 1 to 100) micrograms of CAI active ingredient drug per day, thepatient's disease could be treated for a period ranging from about7-1000 days (1-week to 3-years) via a single administration of thecapsule or device. Such a formulation provides benefits which includeaccurate dosing with heightened patient convenience, because frequentintervention or administration is not required, rather administrationcan be done in some cases only once or twice a decade or even lessfrequently. The formation and loading of microspheres, microcapsules,liposomes, etc. and their ocular implantation are standard techniquesknown by one skilled in the art, for example, the use a ganciclovirsustained-release implant to treat cytomegalovirus retinitis, disclosedin Vitreoretinal Surgical Techniques, Peyman et al., Eds. (MartinDunitz. London 2001, chapter 45); Handbook of Pharmaceutical ControlledRelease Technology, Wise, Ed. (Marcel Dekker, New York 2000), therelevant sections of which are incorporated by reference herein in theirentirety.

B. Dosage Forms

Depending on the clinical needs of a patient, formulations of thesubject invention (see Section IV above) are prepared to either locallydeliver the bioactive CAI immediately after administration or forbioactive CAI release into a target tissue (for example, eye tissue)from its vehicle in a sustained manner over a period of time suitablefor the desired pharmacological action.

An effective quantity of the compound of interest is employed intreatment. The dosage of compounds used in accordance with the inventionvaries depending on the compound and the condition being treated. Forexample, the age, weight, and clinical condition of the recipientpatient; and the experience and judgment of the clinician orpractitioner administering the therapy are among the factors affectingthe selected dosage. Other factors include: the route of administration,the patient, the patient's medical history, the severity of the diseaseprocess, and the potency of the particular compound. The dose should besufficient to ameliorate symptoms or signs of the disease treatedwithout producing unacceptable toxicity to the patient. In general, aneffective amount of the compound is that which provides eithersubjective relief of symptoms or an objectively identifiable improvementas noted by the clinician or other qualified observer. In the case ofangiogenic diseases such as age related macular degeneration anddiabetic retinopathy, the formulations of the present invention areadministered over a course of treatment ranging from weeks to years.

C. Combinatorial Therapies

A further embodiment of the subject invention provides for the localadministration of a CAI compound concurrently with other pharmacologicaltherapies for the treatment of non-life threatening diseases. Forexample, a CAI compound of the invention can be administeredconcurrently with other clinical therapies such as the concurrentadministration with anti-angiogenic compounds (e.g., combretastatin,angiostatin, endostatin, vitaxin, 2ME₂, anecortave, squalamine, macugen,lucentis, PEDF), which may have diverse mechanisms of action (e.g. VEGFneutralization, tyrosine receptor kinase inhibition), glucocorticoidsand non-steroidal anti-inflammatory drugs.

The present invention provides for the concurrent, localizedadministration of therapies and a CAI compound or formulations thereof,wherein the therapy addresses a non-life threatening disease (such ascertain non-cancerous proliferative and angiogenic diseases) other thanthat treated by the CAI compound or formulations thereof, of theinvention. For example, one embodiment of the invention provides thetopical delivery of a CAI prodrug and novel formulations for treatingsevere dermatological diseases including severe psoriasis, excema androsacea and local intrarticular administration for severe arthritis byinhibiting vascular and inflammatory cell proliferation.

A further embodiment of the subject invention provides for the treatmentof neovascular and edematous ocular diseases by ocular administration ofa CAI compound, as described herein, in combination with otherpharmacological anti-angiogenesis therapies. Contemplatedanti-angiogensis therapies comprise those which include, but are notlimited to, glucocorticoids (preferred glucocorticoids include,dexamethasone, fluoromethalone, administration of medrysone, budesonide,betamethasone, fluocinalone, triamcinolone, triamcinolone acetonide,prednisone, prednisolone, hydrocortisone, rimexolone, andpharmaceutically acceptable salts thereof), anecortave acetate,VEGF-binding molecules (oligonucleotide aptamers (e.g. Macugen®),protein antibodies (e.g. Lucentis®), tyrosine receptor kinase inhibitorsincluding but not limited to vascular endothelial growth factor (VEGF),platelet derived growth factor (PDGF) and fibrobrast growth factor (FGF)receptors, other direct or indirect growth factor inhibitors includingsomatostatin receptor agonists (inhibiting release of Growth Hormone andIGF-1), RNAi oligonucleotide transcription inhibitors of ocular diseasemolecular targets including growth factors described above.

Active agents suitable for concurrent administration with a CAI compoundor formulation thereof include, but are not limited to, anti-infectives,including, without limitation, antibiotics, antivirals, and antifungals;antiallergenics and mast cell stabilizers; steroidal and non-steroidalanti-inflammatory agents; cyclooxygenase inhibitors, including, withoutlimitation, Cox I and Cox II inhibitors; combinations of anti-infectiveand anti-inflammatory agents; anti-glaucoma agents, including, withoutlimitation, adrenergics, beta-adrenergic blocking agents, α-adrenergicagonists, parasypathomimetic agents, cholinesterase inhibitors, carbonicanhydrase inhibitors, and prostaglandins; combinations of anti-glaucomaagents; antioxidants; nutritional supplements; drugs for the treatmentof cystoid macular edema including, without limitation, non-steroidalanti-inflammatory agents; drugs for the treatment of ARMD, including,without limitation, angiogenesis inhibitors and nutritional supplements;drugs for the treatment of herpetic infections and CMV ocularinfections; drugs for the treatment of proliferative vitreoretinopathyincluding, without limitation, antimetabolites and fibrinolytics; woundmodulating agents, including, without limitation, growth factors;antimetabolites; neuroprotective drugs, including, without limitation,eliprodil; and angiostatic steroids for the treatment of diseases orconditions of the posterior segment of the eye, including, withoutlimitation, ARMD, CNV, retinopathies, retinitis, uveitis, macular edema,and glaucoma. Such angiostatic steroids are more fully disclosed in U.S.Pat. Nos. 5,679,666 and 5,770,592, which are incorporated herein intheir entirety by reference. Preferred ones of such angiostatic steroidsinclude 4,9(11)-Pregnadien-17α,21-diol-3,20-dione and4,9(11)-Pregnadien-17α,21-diol-3,20-dione-21-acetate. A preferrednon-steroidal anti-inflammatory for the treatment of cystoid macularedema is nepafenac.

A further embodiment of the subject invention provides for treatment ofproliferative and edematous diseases via concurrent administration of aCAI compound and an anti-angiogenic compound. Diverse anti-angiogeniccompounds can be used in a combinatorial treatment of the invention,include those with diverse mechanisms of action (e.g. VEGFneutralization, tyrosine receptor kinase inhibition, arachidonateinhibition, and Bcl-2 upregulation), such as glucocorticoids,non-steroidal anti-inflammatory drugs.

A further embodiment of the subject invention provides for the localadministration of CAI compound and related formulations thereof, incombination with other pharmacological therapies specifically designedto provide efficacy with limited side effects, e.g. neuroprotectantagents, inhibitors of drug efflux, metabolite inhibition, synergisticagents, and agents designed to decrease specific side effects.

A further embodiment of the subject invention provides for the treatmentof ocular diseases such as glaucoma and inflammatory eye disease byocular administration of CAI compound or formulations thereof, asdescribed herein in combination with other pharmacological agentsincluding, but are not limited to, verteporfin photodynamic therapy (QLTPharmaceuticals), anecortave acetate (Alcon Research Ltd.), Macugen(Eyetech Pharmaceuticals), Lucentis (ranibizumab, Genentech), Squalaminelactate (Genaera Corporation), LY333531 (Eli Lilly), and Fluocinolone(Bausch & Lomb), timolol, bromodidine, cyclosporine, cis-platin,carboplatin, methotrexate, steroids, BDNF, CTF, and TNF-α blockers suchas thalidomide and its derivatives and prodrugs.

Dexamethasone has been detected at 13 ng/ml in the vitreous cavityfollowing a single 5 mg peribulbar injection in humans (Weijtens et alOphthalmology 107(10): 1932-8 2000; Weijtens et al Am J Ophthalmol128(2): 192-7 1999). This results in a vitreous concentration ofapproximately 0.10-0.13 μM assuming a vitreous volume of 4-5 ml.Delivery to the subretinal space is 10-fold higher followingsubconjunctival injection (Weijtens et al Am J Ophthalmol 123(3): 358-63(1997)). The serum half-life of dexamethasone (18-36 hours) issignificantly shorter than that of CAI (111 hours). Physiological tissueconcentrations of CAI are in the 1-10 μM range so that it appearsreasonable to postulate based on the scleral permeability and prolongedserum half-life of CAI that it can be effectively deliveredtranssclerally to the subretinal space, and the vitreous cavity inphysiological concentrations (Weijtens et al Ophthalmology 107(10):1932-8 2000; Weijtens et al Am J Ophthalmol 128(2): 192-7 1999). Therelease rates of CAI compound or formulations thereof, from sustainedrelease devices are also favorable. One potential drawback of theintravitreal implants that were constructed with CAI was that somedemonstrated rapid expansion because of the hygroscopic nature of CAI(Weijtens et al Am J Ophthalmol 123(3): 358-63 1997). This rapidexpansion may be alleviated by developing a non-aqueous reservoir forthe CAI compound or formulations thereof, pellet, or by formulations ofa prodrug as stated in this patent.

The present invention also contemplates the use of a glucocorticoidand/or neuroprotective agent in combination with the CAI compound orformulations thereof,. A glucocorticoid alone and/or neuroprotectiveagent in combination with CAI compound or formulations thereof, isuseful for treating persons suffering from pathologic ocularangiogenesis, in particular, exudative AMD and/or PDR, as well assubretinal or retinal edema associated with either condition. Inaddition to being effective in inhibiting the neovascularizationassociated with wet AMD and PDR, CAI compound or formulations thereof,could be useful in controlling any IOP rise associated with the use of aglucocorticoid, or to protect the retina from ischemic damage associatedwith microangiopathy or retinal vascular occlusions.

D. Delivery via Devices

A further embodiment provides the use of CAI compound or formulationsthereof, in conjunction with a drug delivery system in the form of animplant or a device for treatment of conditions as set forth herein.

The present invention also provides CAI compound or formulationsthereof, for use as coatings in conjunction with physical materialimplants such as stents and band ligatures used to treat vasculardiseases. In a preferred embodiment novel CAI compound or formulationsthereof, coated stents of the subject invention are used in thetreatment of vascular disorders such as restenosis or vascular occlusionfollowing vascular insult (e.g., angioplasty, allo- or xenotransplantvasculopathies; variceal bleeding, and transplantation of an organ).

Delivery of drugs in the form of topical eye drops is also of littleutility when the drug is a protein or peptide that lacks the ability tocross the cornea and be made available to the vitreous, retina, or othersubretinal structures such as the retinal pigment epithelium (“RPE”) orchoroidal vasculature. An extraocular insert is a contact lens deliverysystem that releases medication over an extended period. See, e.g.,JAMA, 260:24, p. 3556 (1988). The lens generally only lasts for a matterof hours or days before dissolving or releasing all of the therapeuticcompound. Continuous delivery of medication is inconvenient, requiringfrequent re-application. Again, these contact lenses only provide drugto the cornea and anterior chamber.

In rare cases, direct delivery of drugs has also been accomplished usingextemraliz tubes. This requires insertion of one end of a tube into thecorner of the patient's eye. The other end of the tube is taped to thepatient's forehead and terminates in a septum, through which medicationis delivered. This method is undesirable, being both uncomfortable andinconvenient. Since medication must be injected through the septum, thedevice is incapable of continuous delivery of medication. Furthermore,such tubes may become infected and in some cases ultimately threaten thepatient's vision. Direct delivery of drugs can also be accomplished bythe intraocular injection of the drug, or of microspheres that containor comprise the drug. However, microspheres tend to migrate within theeye, either into the visual axis or into adjacent tissue sites.

An intraocular insert is currently available for delivery of ganciclovirto the eye. Known as Vitrasert, the device consists of a nonerodible,polymer-based, sustained-release package containing ganciclovir, anon-proteinaceous nucleoside analog. The device is surgically implantedin the vitreous humor of the eye to treat cytomegalovirus retinitis.See, e.g., Anand, R., et al., Arch. Ophthalmol., 111, pp. 223-227(1993). Another intraocular insert is disclosed by U.S. Pat. No.5,466,233. This tack-shaped device is surgically implanted so that thehead of the tack is external to the eye, abutting the scleral surface.The post of the tack crosses the sclera and extends into the vitreoushumor, where it provides for vitreal drug release.

CAI compound or formulations thereof, of the invention may beadministered surgically as an ocular implant. As one example, areservoir container having a diffusible wall of polyvinyl alcohol orpolyvinyl acetate or partially hydrolyzed (e.g., about 1% to about 99%hydrolyzed) polyvinyl acetate and containing milligram (e.g., in therange of from about 1 mg to about 100 mg) quantities of CAI may beimplanted in or on the sclera. As another example, CAI in milligramquantities may be incorporated into a polymeric matrix having dimensionsof about 2 mm by 4 mm, and made of a biocompatible and optionallybiodegradable polymer such as polycaprolactone, poly(glycolic) acid,poly(lactic) acid, or a polyanhydride, or a lipid such as sebacic acid,and may be implanted on the sclera or in the eye. This is usuallyaccomplished with the patient receiving either a topical or localanesthetic and using a small (3-4 mm incision) made behind the cornea.The matrix, containing CAI compound or formulations thereof, is theninserted through the incision and sutured to the sclera using 9-0 nylon.

For example, U.S. Pat. No. 5,773,019, the contents of which is herebyincorporated by reference, discloses implantable controlled releasedevices for delivering drugs to the eye wherein the implantable devicehas an inner core containing an effective amount of a low solubilitydrug covered by a non-bioerodible polymer coating layer that ispermeable to the low solubility drug. U.S. Pat. No.5,378,475, thecontents of which is hereby incorporated by reference, disclosessustained release drug delivery devices that have an inner core orreservoir comprising a drug, a first coating layer which is essentiallyimpermeable to the passage of the drug, and a second coating layer whichis permeable to the drug. The first coating layer covers at least aportion of the inner core but at least a small portion of the inner coreis not coated with the first coating layer. The second coating layeressentially completely covers the first coating layer and the uncoatedportion of the inner core. U.S. Pat. No. 4,853,224, the contents ofwhich is hereby incorporated by reference, discloses biodegradableocular implants comprising microencapsulated drugs for implantation intothe anterior and/or posterior chambers of the eye.

The polymeric encapsulating agent or lipid encapsulating agent is theprimary element of the capsule. U.S. Pat. No. 5,164,188, the contents ofwhich is hereby incorporated by reference, discloses the use ofbiodegradable implants in the suprachoroid of an eye. The implants aregenerally encapsulated. The capsule, for the most part, is a polymericencapsulating agent. Material capable of being placed in a given area ofthe suprachoroid without migration, “such as oxycel, gelatin, silicone,etc.” can also be used. U.S. Pat. No.6,120,789, the contents of which ishereby incorporated by reference, discloses the use of a non-polymericcomposition for in situ formation of a solid matrix in an animal, anduse of the composition as a medical device or as a sustained releasedelivery system for a biologically-active agent, among other uses. Suchimplants can provide CAI compound or formulations thereof, of theinvention to a patient to treat a non-life threatening disease.

Another implantable device that can be used to deliver formulations ofthe present invention is the biodegradable implants described in U.S.Pat. No. 5,869,079, the contents of which is hereby incorporated byreference,. Additional intracorporeal devices to which a CAI compound orformulations thereof, can be added for example in the form of a coatinginclude, but are not limited to, catheters, stents, angioplastyballoons, pacemakers, etc.

Within yet other aspects of the present invention, methods are providedfor expanding the lumen of a body passageway, comprising inserting astent into the passageway, the stent having a generally tubularstructure, the surface of the stent structure being coated with acomposition comprising a CAI compound, optionally in the presence ofpaclitaxel, such that the passageway is expanded. Within variousembodiments of the invention, methods are provided for eliminatingbiliary obstructions, comprising inserting a biliary stent into abiliary passageway; for eliminating urethral obstructions, comprisinginserting a urethral stent into a urethra; for eliminating esophagealobstructions, comprising inserting an esophageal stent into anesophagus; and for eliminating tracheal/bronchial obstructions,comprising inserting a tracheal/bronchial stent into the trachea orbronchi. Within each of these embodiments the stent has a generallytubular structure, the surface of the structure being coated with acomposition comprising a CAI compound or formulation thereof and,optionally, together with other pharmaceutically active agents for acombinatin therapy.

V. Animal Efficacy and Safety Models

The CAI parent compound, and the principles on which the CAI compound orformulations thereof, of the inventions described herein derive, havebeen demonstrated to have antiproliferative and antimetastatic activitythat was linked to decrease of intracellular calcium by inhibition ofnon-voltage-gated calcium channels. Furthermore, the anti-angiogenicactivity of CAI, the active ingredient contained in the CAI compound orformulations thereof, or applied via drug delivery systems and routes ofadministration that are the subject of this invention, conferringefficacy on neovascular disease has been well established.

The CAI compound or formulations thereof, used directly or in conjuctionwith a drug delivery method or device as described herein have highlyadvantageous therapeutic efficacy and safety for clinical use as localdisease therapy when compared to systemic therapies of CAI described todate. The advantages of the CAI compound or formulations thereof, andadministration methods of this invention are provided by the controlled,efficacious and safe free CAI drug concentrations to the target tissue.The efficacy of CAI compound or formulations thereof, and drug deliverysystems described herein to treat diseases described herein can beconfirmed using standard in vivo animal test model of disease. Thesemodels include the mouse retinal neovascularization retinopathy ofprematurity model (Smith L E et al. Invest Ophthalmol Vis Sci.35(1):101-11, 1994) the method of which is hereby incorporated byreference as well as the demonstration of an antiproliferative effect onchoroidal endothelial cells and RPE in culture (Hoffman et alOphthalmologe 2004) the method of which is hereby incorporated byreference. Further, the ability of CAI compound or formulations thereof,to provide controlled, safe levels suitable to treat human proliferativeor neovascular ocular diseases can be confirmed using standard in vivoanimal test ocular pharmacokinetic and pharmacodynamic studies inanimals.

The CAI parent compound active ingredient from which the CAI compound orformulations thereof, inventions described herein derive, hasdemonstrated primary pharmacological action to decrease intracellularcalcium concentration by inhibition of non-voltage-gated calciumchannels (Kohn, E. C. et al. CAI. Cancer Res. 54:935-942, 1994) thusaffecting diverse signal transduction processes. Furthermore, thepharmacological mechanisms of action of CAI include (but are not limitedto) the inhibition of disease mediating molecules including growthfactors (e.g. VEGF), cytokines (e.g. IL-6), matrix metalloproteinases(MMps) and arachadonic acid (Felder, C. C et al. J Pharmacol Exp Ther.257:967-971, 1991; Fox D A et al. Ann NY Acad Sci.;893:282-5, 1999; ColeK. et al. Cancer Metastasis Rev. March;13(1):31-44, 1994). Thesepharmacological mechanisms described for CAI provide the fundamentalbasis of anti-proliferative activity on diverse cancer cell types(melanoma, breast, squamous, prostate, ovarian, glioblastoma, colon andsmall lung cell cancer) relevant to current clinical application of CAIas a systemically administered drug in life-threatening cancerindications. Furthermore, these pharmacological mechanisms described forCAI provide the fundamental basis of anti-proliferative activity againstother non-cancer cell types directly relevant to the invention,including but not limited to, human vascular and retinal endothelialcells, retinal pigment and choroidal endothelial cells ((Hoffman et alOphthalmologe 2004), inflammatory cells including lymphocytes andeosinophils. Furthermore, since pathological increases in intracellularcalcium (e.g. as mediated by pathological activation of NMDA receptorchannels by excess glutamate in ischemic conditions) in neurons is knownto result in neuronal death, it is anticipated by this invention thatadministration of CAI compound or formulations thereof, as describedherein can be neuroprotective. Therefore, based on the pharmacologicalmechanisms CAI compound or formulations thereof, described herein haveanti-angiogenic, anti-inflammatory and neuroprotective effectsassociated with the treatment of diseases of relevant diseases asdescribed herein.

The CAI compound or formulations thereof, used directly or in conductionwith a drug delivery method or device as described herein have highlyadvantageous therapeutic efficacy and safety for clinical use as localdisease therapy when compared to systemic therapies of CAI described todate. The advantages of the CAI compound or formulations thereof, andadministration methods of this invention are provided by the controlled,efficacious and safe free CAI drug concentrations administered to orreaching the target tissue. Plasma concentrations of CAI that have beenassociated with efficacy in cancer described in patients range fromapproximately 1.0 to 10 micromolar while plasma concentrations of above100 micromolar can be associated with unacceptable toxicity. Theinvention anticipates pharmacologically active concentrations of CAI asprovided by the CAI compound or formulations thereof, of this inventionto exhibit therapeutic effects in the local target tissue to rangebetween 0.5 and 100 micromolar. As a specific example this inventionanticipates periocular administration of a CAI compound or formulationsthereof, which formulations contain 5 mg of active CAI ingredient, toprovide CAI to the target subretinal and vitreal compartments inconcentrations ranging from 0.1 and 10 micromolar concentration over aone-day time period. The ability of CAI compound or formulationsthereof, to provide controlled, safe levels suitable to diseasesdescribed herein can be confirmed using standard in vivo pharmacokineticstudies in healthy animals or animal disease models. The CAI compound orformulations thereof, is administered as described herein and thesamples of the target tissue are analyzed for concentration of CAIactive principle using known e.g., (Tutsch, K. D. et al. Proc Am AssocCancer Res. 37:A1133, 1996, the contents of which are herebyincorporated by reference in their entirety) or tissue adaptedbioanalytical methods. For example, a CAI formulation containing 5 mg ofactive CAI ingredient is given to rabbits by periocular administration,and the animals are sacrificed at different time points over a 24 hourperiod. Tissue and liquid humor samples are taken from the eyes and theCAI active ingredient is extracted and subjected to HPLC-MS analysiswith an appropriate internal standard to the target disease compartmentsincluding retinal, choroidal and vitreal compartments.

The efficacy of CAI compound or formulations thereof, alone or inconjunction with drug delivery systems described herein to treatdiseases described herein can be confirmed using in vivo animal testmodels of disease. These models include the mouse retinalneovascularization retinopathy of prematurity model (Smith L E et al.Invest Ophthalmol Vis Sci. 35(1):101-1 1, 1994, the methods of which arehereby incorporated by reference). In this model, mouse pups are placedinto 75% O₂, along with their nursing dams, at post-natal day 7. Theyare maintained at this oxygen level for five days, at which point theyare returned to normoxia. For the next five days, the pups areadministered with CAI compound or formulations thereof, using themethods described herein. At day 17, the animals are euthanized,enucleated, and the eyes fixed in 4% buffered paraformaldehydeovernight, then transferred to saline, embedded in paraffin, andsectioned. Serial sections are collected (6 micron sections, every30^(th) section) and stained with hematoxylin and eosin. Individualsmasked to the identity of treatment then count the number of pre-retinalnuclei and the effects of CAI compound or formulations thereof, onlowering the number of pre-retinal nuclei are evaluated relative tocontrol. At least eight sections are counted for each eye. Another modelis the adult mouse model of ischemia-induced choroidalneovascularization that mimics age-related macular degeneration. In thismodel adult mice are subjected to laser photocoagulation in a mannersimilar to Ryan's (Ryan S J. Et al. Trans Am Ophthalmol Soc. 77:707-745,1979, the methods of which are hereby incorporated by reference). Threeburn spots are produced in the choroid using an argon green wavelengthlaser at a power of 910-1030 mW for 0.05 sec to induce choroidal ruptureand subsequent neovascularization. The burns are in three quadrants ofthe choroid, one disc area in diameter and one disc area from the opticnerve. This fixed distance allows the burns to be both reproducible andisolated from each other. The laser produces a bubble in the majority ofcases, which is indicative of a Bruch's membrane rupture. In less than5% of the animals, bleeding may be noted upon treatment. The mice areeuthanized at two weeks after receiving laser photocoagulation and theeyes removed for evaluation of choroidal neovascularization.

Other disease models include xenograft models for host vs. graft tissuerejection relevant to applications including restenosis. Another objectof the present invention is to provide for local treatment of otherproliferative and angiogenic diseases including topical delivery of CAIcompound or formulations thereof, for severe dermatological diseasesincluding severe psoriasis, excema and rosacea and local intrarticularadministration for severe arthritis by inhibiting vascular andinflammatory cell proliferation.

VI. Therapeutic Applications

In one embodiment, the CAI compound and formulations thereof of thepresent invention can be used for administration to a human patientsuffering from a non-life threatening disease, as a method of treatmentof such disease, which non-life threatening disease can be, for examplea proliferative disease, an inflammatory disease, an edematous disease,a neurodegenerative and/or a neurotoxic disease, or a signaltransduction-mediated disease, or a matrix matalloproteinase-mediateddisease.

Examples of diseases or disorders that can be mediated directly, orindirectly, by the administration of a CAI compound or formulationsthereof, of the invention include, but are not limited to: age-relatedmacular degeneration, diabetic retinopathy, retinal vascular occlusion,choroidal and retinal angiomatous proliferation, chronic glaucoma,retinal detachment, sickle cell retinopathy, rubeosis iritis, uveitis,neoplasms, Fuch's heterochromic iridocyclitis, neovascular glaucoma,corneal neovascularization, neovascularization resulting from combinedvitrectomy and lensectomy, retinal ischemia, choroidal vascularinsufficiency, choroidal thrombosis, carotid artery ischemia, retinalartery/vein occlusion, e.g., central retinal artery occlusion and branchretinal vein occlusion, contusive ocular injury, and retinopathy ofprematurity, and other vascular anomalies, e.g., retinitis pigmentosa,endophthalmitis, infectious diseases, inflammatory but non-infectiousdiseases, ocular ischemia syndrome, peripheral retinal degenerations,retinal degenerations and tumors, choroidal disorders and tumors,vitreous disorders, retinal detachment, non-penetrating and penetratingtrauma, post-cataract complications, and inflammatory opticneuropathies.

Hereditary degenerative retinal and vitreoretinal diseases treatablewith a CAI compound or formulations thereof, of the invention, eitheralone or in combination therapies, include: Primary pigmentedretinopathies, all gene types (ocular involvement only); Autosomaldominant retinitis pigmentosa e.g. rod-cone and cone-rod degenerations;Autosomal recessive retinitis pigmentosa e.g. rod-cone and cone-rodedegenerations, Leber's amaurosis congenita; X-linked recessive pigmentedretinopathies e.g. choroideremia. Secondary pigmented retinopathies(retinopathies associated with systemic diseases); Autosomal dominantpigmented retinopathies, e.g. Paget's disease, Charcot-Marie-Toothdisease, Steinert's disease, Pierre-Marie syndrome; Autosomal dominantpigmented retinopathies e.g. diabetes mellitus, mannosidoses,mucopolysccharidoses, Batten's disease, Refsum's disease, Ushersyndrome; X-linked recessive pigmented retinopathies e.g. Huntersyndrome; conjunctivitis (e.g. allergic conjunctivitis, chronicconjunctivitis, contact lens-associated conjunctivitis, conjunctivalulceration, drug-related conjunctivitis); uveitis, uveoretinitis,chronic diseases (e.g. age-related macular panuveitis, retinitis,degeneration diabetes mellitus, infectious choroiditis, vitreitis,diseases (e.g., tuberculosis syphilis,cytomegalo-Scleritis/Episcleritis, virus retinitis), injury as a resultof physical agents (e.g. UV Iridocyclitis, radiation), chemical agents(e.g. acids, Endophthalmitis caustic solvents), and immunologicaletiologies (e.g. sarcoidosis, inflammatory bowel disease, Cornealulceration disease, and other collagen vascular diseases). VonHippel-Lindau syndrome is a specific neovascular disease that has bothocular and non-ocular manifestations that should be treatable by CAIcompounds and formulations thereof.

The compounds, formulations, and methods of the present invention mayalso be used to treat ocular symptoms resulting from diseases orconditions that have both ocular and non-ocular symptoms. Some examplesinclude AIDS-related disorders such as cytomegalovirus retinitis anddisorders of the vitreous; pregnancy-related disorders such ashypertensive changes in the retina; and ocular effects of variousinfectious diseases such as tuberculosis, syphilis, lyme disease,parasitic disease, toxocara canis, ophthalmonyiasis, cyst cercosis, andfungal infections. Non ocular diseases can also include rheumatoidarthritis, psoriasis, contact dermatitis, keratitis, conjunctivitis,scleritis, squamous cell carcinoma and condyloma.

Angiogenesis and neovascularization in the adult animal is usually apathological process, and is in direct contradistinction tonon-pathological neovascularization, which usually occurs in normalembryogenesis (e.g., development of the embryonic vascular system). Inaccordance with the subject invention, neovascularization refersspecifically to pathological neovascularization. Aberrant orpathological vascularization is a key component in numerous diseasestates. For example, vascularization is a critical element of most solidtumors, such as cancers of the brain, genitourinary tract, lymphaticsystem, stomach, larynx and lung. These include histiocytic lymphoma,lung adenocarcinoma and small cell lung cancers. Aberrant vasculargrowth in the retina can lead to visual degeneration which can culminatein blindness. Accordingly, the subject invention provides CAI compoundand formulations thereof for the treatment of neovascularization.

Compounds of the invention can also be used to inhibit the proliferationof vascular endothelial cells and so are indicated for use in treatinggraft vessel diseases such as restenosis or vascular occlusion followingvascular insult such as angioplasty, allo- or xenotransplantvasculopathies, graft vessel atherosclerosis, and in the transplantationof an organ (e.g., heart, liver, lung, kidney or pancreatic transplants(Weckbecker et al., Transplantation Proceedings 1997, 29, 2599-2600).

The present invention also provides for the treatment of severedermatological diseases including severe psoriasis, contact dermatitis,excema and rosacea; severe arthritis; and other vascular andinflammatory cell proliferative diseases (such as rheumatoid arthritis;spondyloarthropathies; gouty arthritis; osteoarthritis; systemic lupuserythematosus; juvenile arthritis; asthma; bronchitis; menstrual cramps;tendonitis; bursitis; skin related conditions such as psoriasis, eczema,burns, and dermatitis; gastrointestinal conditions such as infammatorybowel disease, Crohn's disease, gastritis, irritable bowel syndrome andulcerative colitis; colorectal cancer; migraine headaches; periarteritisnodosa; thyroiditis; aplastic anemia; Hodgkin's disease; sclerodoma;rheumatic fever; type I diabetes; myasthenia gravis; sarcoidosis;nephrotic syndrome; Behcet's syndrome; polymyositis; gingivitis;hypersensitivity; swelling occurring after injury; myocardial ischemia;and the like).

Additionally, disease states which rely on aberrant signaltransduction/proliferation may also be treated by the CAI compound orformulations thereof, or methods of the invention. Diseases ofpotentially aberrant signal transduction/proliferation may include thecollagen vasculitides (i.e., systemic lupus erthythematosis andrheumatoid arthritis), neurologic diseases (i.e., dementia and nerveconduction diseases), diseases of transport (i.e., cystic fibrosis),toxic effects of agents (i.e., cisplatin-related neuropathy), andcellular dysfunction (i.e., myelodysfunction syndromes), hemangiomata,and collagen vasculidities.

One embodiment of the subject invention provides for the local ocularadministration of CAI compound or formulations thereof, by means ofperiocular, retrobulbar, intravitreal, subretinal, posteriorjuxtascleral, topical and subconjunctival administration throughinjection or needle-free system, or topical administration though localinstillation, drops, ointment, or in conjunction with drug deliverysystems exemplified by contact lenses, devices and implants. A relatedembodiment provides the treatment of a broad range of ocular diseasesincluding diabetic retinopathy (DR), neovascular proliferativeage-related macular degeneration (ARMD), diabetic macular edema (DME),cystoid macular edema (CME) and ocular tumors such as retinoblastoma(RB), Retinopathy of Prematurity (ROP), retinal vascular occlusions(RVO), uveitis, glaucoma, corneal neovascularization, irisneovascularization, neovascular glaucoma, ischemic neural damage, andpterygium by the local ocular administration of CAI compound and relatedformulations therof, directly or in conjuction with a drug deliverysystem as described herein.

Following are examples, which illustrate procedures for practicing theinvention. These examples should not be construed as limiting.

It is advantageous to define several abbreviations before describing theexamles. It should be appreciated that the following abbreviations areused throughout this application.

-   CAI=5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide-   Tween 80=polyoxyethylenesorbitan monooleate-   Pluronic=polyoxyethylene-polyoxypropylene block copolymer-   Pluronic F-68,    F-127=α-hydro-omega-hydroxypoly(oxyethylene)-poly(oxypropylene)-poloxyethylene    block copolymers

EXAMPLE 1 Procedures for Preparing and Identifying Stable, AutoclavableAqueous Formulations of CAI Compounds

General Procedure

Amorphous solid free base form of CAI active pharmaceutical ingredienthaving Formula I (5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4-carboxamide), which can be synthesized by one skilled inthe art of organic chemistry according to procedures described in U.S.Pat. Nos. 4,590,201 and 5,602,156, is first dissolved in a minimalvolume of absolute ethanol at 40° C. Additives are added to thissolution as selected from an ionic surfactant (preferably a lipophilicorganic acid acid having at least 8 carbon atoms such as oleic acid), anorganic oil, such as alkyl ester of fatty acid and/or monoglyceride,diglyceride, or triglyceride, or glyceride (e.g. soybean oil, ormixtures thereof), and other organic solubilizing additives (e.g.Tweens, Pluronics, cyclodextrins, polysorbates, sorbitan esters,sucroesters, and other non-ionic surfactants, polyethylene glycols,particularly those that are liquid at ambient temperature) inproportions that result in a clear solution containing CAI inconcentrations from 1 to 200 mg/mL. Preferred proportions of theseprimary formulation ingredients are selected based on suitability forachieving concentrations of CAI from 1 to 100 mg/mL. After adding waterto the resulting organic solution of CAI a stable organic-aqueousmixture comprising CAI results as a homogeneous solution, stableemulsion, or self-emulsifying dry powder. The ethanol is removed fromthe resulting organic mixture described above by evaporation (e.g. underreduced pressure using a rotary evaporator). Water is added to theresulting organic mixture, and the pH is adjusted to an acceptabletherapeutically applicable range (preferably from about pH 3.0 to aboutpH 8.0 and more preferably from about pH 5.0 to about pH 8.0). The exactpH adjustment is done by addition of a basic solution (preferablyaqueous sodium hydroxide) or an acidic solution (preferably aqueoushydrochloric acid) or both in quantities sufficient to provide thedesited pH.

Preliminary preferred formulations are first selected by simpleinspection based on stability of the final solution against separationinto distinct organic and aqueous phases after standing for at least 3days within an ambient temperature range. Formulations that phaseseparate are unstable and are discarded. Stable sterile formulations arethen prepared by autoclaving for sufficient time at temperaturessuitable for effectively killing the microorganisms that may be presentin the formulation, for instance, 121° C for about 30 minutes.

An alternate method for manufacturing stable sterile formulations can bereadily accomplished by filtration through sterile filtration membranes(e.g., 0.1 micron membranes, 0.2 micron membranes).

Formulation 1

Formulation 1 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (150 mg),soybean oil (150 mg), Tween-80 (100 mg), PluronicF-68 (100 mg). A creamyemulsion of milky white appearance is formed after dilution with waterto a final CAI concentration of 36 mg/mL (3.6%). The emulsion ofFormulation 1 is stable based on stability to phase separation asdefined in Example 1. The active CAI ingredient is stable to autoclavesterilization conditions of 121° C. at a pressure of 15 psi as definedin Example 1. Thus, after autoclaving Formulation 1 under theseconditions for 30 min no decomposition of CAI or formation of newimpurities could be detected by reverse phase high pressure liquidchromoatography (RP-HPLC) analysis using UV detection (Conditions: RP-18column: 5μ particle size, length=13 cm, diameter=8 mm; solvent system:solvent A=0.001% trifluoroacetic acid, solvent B=acetonitrile,gradient=80% A/20% B to 10% A/90% B over 12 min at a flow rate of 2.5mL/min; CAI retention time=6.3 min).

Formulation 2

Formulation 2 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (200 mg),soybean oil (200 mg) and Tween-80 (100 mg). An emulsion of milky whiteappearance is formed after dilution with water to a final CAIconcentration of 60 mg/mL (6.0%). The emulsion of Formulation 2 isstable based on stability to phase separation at ambient temperature.The active CAI ingredient is stable to autoclave sterilizationconditions of 121° C. at a pressure of 15 psi as defined in Example 1.Thus, after autoclaving Formulation 2 under these conditions for 30 minno decomposition of CAI or formation of new impurities could be detectedby RP-HPLC analysis using UV detection (Conditions: RP-18 column: 5 pparticle size, length=13 cm, diameter=8 mm; solvent system: solventA=0.001% trifluoroacetic acid, solvent B=acetonitrile, gradient=80%A/20% B to 10% A/90% B over 12 min at a flow rate of 2.5 mL/min; CAIretention time=6.3 min).

Formulation 3

Formulation 3 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (300 mg),Pluronic F-127 (100 mg) and Tween-80 (100 mg). An creamy emulsion ofmilky white appearance is formed after dilution with water to a finalCAI concentration of 36 mg/mL (3.6%). The emulsion of Formulation 3 isstable based on stability to phase separation at ambient temperature.The active CAI ingredient is stable to autoclave sterilizationconditions of 121° C. at a pressure of 15 psi as defined in Example 1.Thus, after autoclaving Formulation 3 under these conditions for 30 minno decomposition of CAI or formation of new impurities could be detectedby RP-HPLC analysis using UV detection (Conditions: RP-18 column: 5μparticle size, length=13 cm, diameter=8 mm; solvent system: solventA=0.001% trifluoroacetic acid, solvent B=acetonitrile, gradient=80%A/20% B to 10% A/90% B over 12 min at a flow rate of 2.5 mL/min; CAIretention time=6.3 min).

Formulation 4

Formulation 4 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (150 mg),soybean oil (150 mg), Pluronic F-68 (100 mg) and Tween-80 (100 mg). Acreamy emulsion of milky white appearance is formed after dilution withwater to a final CAI concentration of 36 mg/mL (3.6%). The emulsion ofFormulation 4 is stable based on stability to phase separation atambient temperature. The active CAI ingredient is stable to autoclavesterilization conditions of 121° C. at a pressure of 15 psi. as definedin Example 1. Thus, after autoclaving Formulation 4 under theseconditions for up to 30 min no decomposition of CAI or formation of newimpurities could be detected by RP-HPLC analysis using UV detection(Conditions: RP-18 column: 5μ particle size, length=13 cm, diameter=8mm; solvent system: solvent A=0.001% trifluoroacetic acid , solventB=acetonitrile, gradient=80% A/20% B to 10% A/90% B over 12 min at aflow rate of 2.5 mL/min; CAI retention time=6.3 min).

Formulation 5

Formulation 5 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (50 mg), oleic acid (150 mg),soybean oil (150 mg), Pluronic F-127 (100 mg) and Tween-80 (100 mg). Acreamy emulsion of milky white appearance is formed after dilution withwater to a final CAI concentration of 36 mg/mL (3.6%). The emulsion ofFormulation 4 is stable based on stability to phase separation atambient temperature. The active CAI ingredient is stable to autoclavesterilization conditions of 121° C. at a pressure of 15 psi as definedin Example 1. Thus, after autoclaving Formulation 5 under theseconditions for 30 min no decomposition of CAI or formation of newimpurities could be detected by RP-HPLC analysis using UV detection(Conditions: RP-18 column: 5μ particle size, length=13 cm, diameter=8mm; solvent system: solvent A=0.001% trifluoroacetic acid , solventB=acetonitrile, gradient=80% A/20% B to 10% A/90% B over 12 min at aflow rate of 2.5 mL/min; CAI retention time=6.3 min).

Formulation 6

Formulation 6 is prepared according to the General Procedure describedin Example 1 using the following amounts of reagents or correspondingrelative proportions: CAI free base (25 mg), oleic acid (150 mg),soybean oil (150 mg), Pluronic F-68 (100 mg) and Tween-80 (100 mg). Ancreamy emulsion of milky white appearance is formed after dilution withwater to a final CAI concentration of 8 mg/mL (0.8%). The emulsion ofFormulation 6 is stable based on stability to phase separation atambient temperature. The active CAI ingredient is stable to autoclavesterilization conditions of 121° C. at a pressure of 15 psi as definedin Example 1. Thus, after autoclaving Formulation 6 under theseconditions for 30 min no decomposition of CAI or formation of newimpurities could be detected by RP-HPLC analysis using UV detection(Conditions: RP-18 column: 5μ particle size, length=13 cm, diameter=8mm; solvent system: solvent A=0.001% trifluoroacetic acid , solventB=acetonitrile, gradient=80% A/20% B to 10% A/90% B over 12 min at aflow rate of 2.5 mL/min; CAI retention time=6.3 min).

EXAMPLE 2 Intravitreal Injection

Pharmacologically effective concentrations in diverse in vitro and invivo model systems of CAI at 0.5 to 10 micromolar concentration which isconsistent with a dose of 0.2 ng to 40 ng of drug in a compartment ofabout 4 to 5 mL for the eye for a long term 4 to 6 week maintenance of acontrolled dose use with indicate a dose range of 50 ng to 500 ng of CAIactive ingredient or greater.

The intravitreal injection would be administered via the inferotemporalpars plana, in a protocol similar to triamcinalone, Macugen, orLucentis. As a single dose of 5 mg of dexamethasone administeredsubconjunctivally can reach micromolar concentrations in the subretinalspace and vitreous, and CAI possesses similar trans-scleral permeabilitykinetics, we would propose an initial dose of 5 to 50 mg for periocularadministration. An effective quantity of the compound of interest isemployed in treatment.

As understood by the skilled clinician, the dosage of compounds used inaccordance with the invention varies depending on the compound and thecondition being treated. For example, the age, weight, and clinicalcondition of the recipient patient; and the experience and judgment ofthe clinician or practitioner administering the therapy are among thefactors affecting the selected dosage. Other factors include: the routeof administration, the patient, the patient's medical history, theseverity of the disease process, and the potency of the particularcompound. The dose should be sufficient to ameliorate symptoms or signsof the disease treated without producing unacceptable toxicity to thepatient. In general, an effective amount of the compound is that whichprovides either subjective relief of symptoms or an objectivelyidentifiable improvement as noted by the clinician or other qualifiedobserver.

EXAMPLE 3 Single Ingredient Formulation

Ingredient Concentration w/v % CAI 0.1 to 2% Monobasic Sodium PhosphateDihydrate 0.051% Dibasic Sodium Phosphate Dodecahydrate 0.5% Tyloxapol0.05-0.4% Sodium Chloride 0.76% NaOH/HCl pH adjust to 5.0-8.4 Water forinjection q.s. 100%.

EXAMPLE 4 Steroid Combination Formulation

Ingredient Concentration w/v % CAI 0.1 to 2% Triamcinolone Acetonide0.5-4.0% Monobasic Sodium Phosphate Dihydrate 0.051% Dibasic SodiumPhosphate Dodecahydrate 0.5% Tyloxapol 0.05-0.4% Sodium Chloride 0.76%NaOH/HCI pH adjust to 5.0-8.4 Water for injection q.s. 100%.

EXAMPLE 5 Neuroprotective Combination Formulation

Ingredient Concentration w/v % CAI 0.1 to 2% Neuroprotecive agent0.5-4.0% Monobasic Sodium Phosphate Dihydrate 0.051% Dibasic SodiumPhosphate Dodecahydrate 0.5% Tyloxapol 0.05-0.4% Sodium Chloride 0.76%NaOH/HCI pH adjust to 5.0-8.4 Water for injection q.s. 100%.

EXAMPLE 6 Chelating Agent Combination Formulation

Ingredient Concentration w/v % CA 0.1 to 2% Chelating Agent 0.5-4.0%Monobasic Sodium Phosphate Dihydrate 0.051% Dibasic Sodium PhosphateDodecahydrate 0.5% Tyloxapol 0.05-0.4% Sodium Chloride 0.76% NaOH/HCI pHadjust to 5.0-8.4 Water for injection q.s. 100%.

EXAMPLE 7 Topical Formulation

A typical example of topical formulation of CAI is as follows:Ingredient Concentration w/v % (Preferred Range) CAI Polyquad0.0005-0.01% (0.0001%) HPMC (hydroxypropylmethylcellullose) 0.02-1.0%(0.5%) Mannitol (b) 0.0-5.0% (3.82%) Sodium Chloride (d) 0.0-0.8%(0.17%) Disodium Edetate 0.0-0.2% (0.01%) Polysorbate-80 (c) 0.005-0.4%(0.05%) NaOH and/or HCl q.s. pH 5.0-8.4 (6.8-7.8) Purified Water q.s.100% (a) other suitable polymers include cellulosic polymers like HPMC,HEC (hydroxyethylcellulose), sodium CMC (sodiumcarboxymethylcellulose)), polyvinyl alcohol (PVA), Polyvinyl Pyrrolidone(PVP), polyacrylamide, and other water miscible/soluble polymers toimpart viscosity to the product and to stabilize suspension. (b) bothionic as well nonionic agents are used to adjust Osmolality of theproduct either alone or in combination. This also stabilizes thesuspension. (c) other surfactants that can be used are non-ionic(Tyloxapol, Tweens, Spans) anionic (lecithin, hydrogenated lecithins),or anionic (sodium lauryl sulfate, bile salts).

The following contemplated clinical cases of angiogenesis are presentedas examples of methods of treatment and not as limitations. Otherillustrations of treating systemic or localized angiogenesis are notpresented, since the principles involved are similar, and will beapparent to the skilled reader based upon review of an individual'sclinical manifestation and the preferred embodiments of the inventivecompositions.

EXAMPEL 8 Diabetic Complications

Patient A is determined to be experiencing an acute relapse of diabetes,e.g., diabetic coma. Patient A has a history of diabetes over the past20 years, with increasing debilitation from progressive diabeticretinopathy, nephropathy, and neuropathy. Patient A has not yet hadpanretinal laser photocoagulopathy but has been treated with focal laserfor macular edema. Retinal examination reveals Patient A has notprogressed to threshold criteria for panretinal photocoagulation per theDiabetic Retinopathy Study (DRS) guidelines.

According to the subject invention, Patient A will require scheduling ofa follow-up visit and low-dose treatment with a CAI compound of theinvention in an attempt to delay or remove the need for panretinal laserphotocoagulopathy. Patient A is given a direct intravitreal injection ofa high dose of CAI, 50 microgram/injection every week for 4 weeks. Thedose remains constant with weekly injections and Patient A is scheduledfor panretinal laser photocoagulopathy with continued weekly retinalexamination follow-up to determine disease course and confirm necessityfor panretinal laser photocoagulopathy.

After regression of the neovascularization, Patient A is treated withfocal laser therapy for macular edema, if needed, and provided a withmonthly, or less frequent, intravitreal injections of CAI with theadministration frequency and dose adjusted according to patientsclinical manifestations and to maintain neovascular regression.Intravitreal injections may be supplemented or replaced by otherappropriate routes of administration and this could be dictated by thepatients clinical presentation, health, expected treatment complianceprofile, and appropriateness of local, systemic, or controlled releasetreatment. CAI treatment would be targeted to be tapered off toprophylactic systemic or controlled release therapy, as the conditionimproves sufficiently.

Patient B is also determined to be experiencing an acute relapse ofdiabetes, e.g., diabetic coma and has a history of diabetes over thepast 20 years, with increasing debilitation from progressive diabeticretinopathy, nephropathy, and neuropathy. Patient B has not yet hadpanretinal laser photocoagulopathy nor has she been treated with focallaser for macular edema. Patient B has not developed thresholdproliferative diabetic retinopathy. This patient is clinically similarto Patient A, but the evidence of no macula involvement obviates theneed for scheduling focal laser photocoagulation.

According to the subject invention, Patient B is given a directintravitreal injection of a low dose of CAI, 5 microgram/injection. Thedose remains constant with a second intravitreal injection 4 weekslater. Patient B is followed clinically every 3 months and treated withfocal laser therapy, if needed, or provided with subsequent intravitrealinjections of CAI with the administration frequency and dose adjustedaccording to Patient B's clinical manifestations. Intravitrealinjections may be supplemented or replaced by other appropriate routesof administration and this could be dictated by the patients clinicalpresentation, health, expected treatment compliance profile, andappropriateness of local, systemic, or controlled release treatment. CAItreatment is targeted as prophylactic systemic or controlled releasetherapy, if the condition improves sufficiently.

EXAMPLE 9 Proliferative Diabetic Retinopathy

62-year old male with Proliferative Diabetic Retinopathy and thresholdhigh-risk characteristics presents with Vitreous Hemorrhage. The patientalso has atrial fibrillation and ischemic cardiomyopathy that requiresoral Coumadin. The patient is intolerant to topical PanretinalPhotocoagulation. The INR measured at just less than 2. Because of thisacute event intravitreal injection, a medium dose of 50 mcg CAI isadministered with some neovascular regression. The patients VitreousHemorrhage slowly resolves and the patient is able to taper his Coumadinso that Panretinal Photocoagulation can be placed with retrobulbaranesthesia. Alternately the patient remains on Coumadin and then istreated with a patch periocular insert of CAI to diminish theneovascularization.

EXAMPLE 10 Central Scotoma Treatment Complications

78-year old woman with a two-week history of central scotoma. Shedeveloped a large disciform scar in the contralateral eye after severalsessions of photodynamic therapy. On exam and fluorescein angiography ispredominately classic subfoveal membrane is noted. Intravitrealinjection of a high dose, 200 mcg, CAI is given to hasten theneovascular regression. After this high dose of intravitreal CAI aperiocular injection of CA in a sustained release solution is given andneovascular regression occurs.

EXAMPLE 11 Age-Related Macular Degeneration Complication

67-year old white male who has had three years of poor vision secondaryto a disciform scar associated with Age-Related Macular Degeneration. Hehas high-risk drusen and parafoveal pigmentation. A long acting low dosesubconjunctival or periocular implant of CAI 1 mg is affixed to thewell-sighted eye to prophylax against neovascularization.

EXAMPLE 12 Glaucoma Complication

82-year old woman with low-tension glaucoma. She has a history ofhypotony maculopathy in the left eye after trabeculectomy with visualacuity loss to 20/200 secondary to hypotony maculopathy associated withan intraocular pressure of 7 to 8. In the right eye she is progressivelycupping with a Cup/Disc ratio 0.85 despite maximal medical treatment andan intraocular pressure of 11. A low dose 1 mg periocular insert forinjection of CAI is given in order to maximize neuroprotection.

EXAMPLE 13 Proliferative Diabetic Retinopathy

28-year old noncompliant diabetic male who initially presented withaggressive Proliferative Diabetic Retinopathy. Heavy and full PanretinalPhotocoagulation was performed, however there is recurrent residualVitreous Hemorrhaging from a small active frond of neovascularizationthat is within heavy Panretinal Photocoagulation scars. An intravitrealinjection of moderate dose, 100 mcg of CAI is given to cause neovascularinvolution and the patient is maintained with a moderate dose of 10 mgadministered by periocular injection of CAI in a sustained releasesolution.

EXAMPLE 14 Proliferative Diabetic Retinopathy

37-year old white male who is with Proliferative Diabetic Retinopathyand has undergone Trans Pars Plana Vitrectomy ×'s 2 for VitreousHemorrhage. He has a heavy pattern of Panretinal Photocoagulation scarsand it is thought that he has neovascularization arising from thesclerotomy sites, and/or the pars plana which is causing persistenthemorrhage. A high dose of 200-mcg dose of CAI is administeredintravitreally and then two weeks later an outpatient air-fluid exchangeis performed, and a moderate dose, periocular injection of 10 mg of CAIis given to prevent further Vitreous Hemorrhage.

EXAMPLE 15 Vision Loss

16-year old female who presents with a several month history of visionloss and is found to have a large angiomatous lesion that is causing asubtotal retinal detachment. Secondary to the lesion size it is notamenable for cryotherapy and standard laser photocoagulation. Initiallya high dose of 200 mcg CAI is given intravitreally and two weeks later aperiocular insert is placed of 10 mg, moderate dose of CAI onto thesclera overlying the lesion.

EXAMPLE 16 Central Retinal Vein Occlusion

66-year old male who presents with a Central Retinal Vein Occlusion andflorid Iris Neovascularization with Neovascular Glaucoma. As an adjunctto Panretinal Photocoagulation secondary to the florid neovascularactivity, a high dose of 200 mcg CAI is given as a synergistic measureto cause regression of the anterior segment neovascularization andnormalization of the pressure.

EXAMPLE 17 Iris and Preretinal Neovascularization

39-year old white male strikes his head on a laboratory cabinet. Thiscauses a chronic peripheral retinal detachment with irisneovascularization and preretinal neovascularization. In order todecrease the risk of intraocular hemorrhage and glaucoma associated withretinal reattachment, a moderate dose of 100 mcg of CAI is givenintravitreally and one-week later successful retinal reattachmentsurgery is performed.

EXAMPLE 18 Proliferative Sickle Cell Retinopathy

40-year old male with a history of Traction Retinal Detachment andvision loss in one eye from proliferative sickle cell retinopathy. Thepatient developed a Vitreous Hemorrhage in the fellow and has onlyincomplete regression following peripheral scatter laser. Relatively lowdose CAI 1 mg is administered periocularly to cause involution of thesmall amount of residual neovascularization which is causing persistentVitreous Hemorrhage.

EXAMPLE 19 Choroidal Neovascular Membrane Complications

44-year old white male with a history of Choroidal Neovascular Membranesecondary to high myopia. One eye has Fuch's spots and 20/200 vision.The second eye is affected with a subfoveal Choroidal NeovascularMembrane and 20/80 vision. A single injection of moderate dose, 100 mcg,of CAI is administered intravitreally to hasten neovascular regression.

EXAMPLE 20 Photodynamic Therapy Complications

32-year old female with multifocal choroiditis and macular scarring inone eye after several sessions of Photodynamic Therapy. She presents inthe second eye with decreased vision and a subfoveal ChoroidalNeovascular Membrane. A mixture of 4 mg of Kenalog, and a high dose of200 mcg of intravitreal CAI are administered to hasten neovascularregression.

It should be understood that the embodiments of the invention shown anddescribed in the specification are only preferred embodiments of theinventor who is skilled in the art and are not limiting in any way.Therefore, various changes, modifications or alterations to theseembodiments may be made or resorted to without departing from the spiritof the invention and the scope of the following claims.

1. A method for treating a patient suffering from an ocular disease orsymptom comprising: diagnosing an ocular disease or symptom in apatient; local ocularly administering to said patient a sterile, aqueousformulation comprising a therapeutically effective amount of a CAIcompound.
 2. The method of claim 1, wherein the CAI compound is selectedfrom the group consisting of CAI free base; CAI acid addition salt, andCAI prodrug.
 3. The method of claim 1, wherein said ocular disease orsymptom is selected from the group consisting of: age-related maculardegeneration, diabetic retinopathy, retinal vascular occlusion,choroidal and retinal angiomatous proliferation, chronic glaucoma,retinal detachment, sickle cell retinopathy, rubeosis iritis, uveitis,neoplasms, Fuch's heterochromic iridocyclitis, neovascular glaucoma,corneal neovascularization, neovascularization resulting from combinedvitrectomy and lensectomy, retinal ischemia, choroidal vascularinsufficiency, choroidal thrombosis, carotid artery ischemia, retinalartery/vein occlusion, contusive ocular injury, retinopathy ofprematurity, retinitis pigmentosa, endophthalmitis, infectious diseases,inflammatory but non-infectious diseases, ocular ischemia syndrome,peripheral retinal degenerations, retinal degenerations and tumors,choroidal disorders and tumors, vitreous disorders, retinal detachment,non-penetrating and penetrating trauma, post-cataract complications,inflammatory optic neuropathies, hereditary degenerative retinal andvitreoretinal diseases, primary pigmented retinopathies, Autosomaldominant retinitis pigmentosa, Auto somal recessive retinitispigmentosa, Leber's amaurosis congenital, X-linked recessive pigmentedretinopathies, secondary pigmented retinopathies, autosomal dominantpigmented retinopathies, Paget's disease, Charcot-Marie-Tooth disease,Steinert's disease, Pierre-Marie syndrome, Autosomal dominant pigmentedretinopathies, mannosidoses, mucopolysccharidoses, Batten's disease,Refsum's disease, Usher syndrome, X-linked recessive pigmentedretinopathies, Hunter syndrome, conjunctivitis, allergic conjunctivitis,chronic conjunctivitis, contact lens-associated conjunctivitis,conjunctival ulceration, drug-related conjunctivitis, uveitis,uveoretinitis, age-related macular panuveitis, retinitis, degenerationdiabetes mellitus, infectious choroiditis, vitreitis, tuberculosissyphilis, cytomegalo-Scleritis/Episcleritis, virus retinitis, UVIridocyclitis, sarcoidosis, and inflammatory bowel disease, cornealulceration disease, and von Hippel-Lindau syndrome.
 4. The method ofclaim 1, wherein the local ocular administration is by topicaladministration or ocular injection.
 5. The method of claim 4, whereinthe ocular injection is any one or combination of routes selected fromthe group consisting of periocular injection, subtenon's injection,juxtascleral injection, intravitreal injection, subconjuctivalinjection, subretinal injection, and retrobulbar injection.
 6. Themethod of claim 1, wherein the therapeutically effective amount of theCAI compound is from 0.1 mg/mL to 100 mg/mL of formulation.
 7. Themethod of claim 6, wherein the therapeutically effective amount of theCAI compound is from 1 mg/mL to 60 mg/mL of formulation.
 8. The methodof claim 1, wherein the CAI compound is a solid particle suspended inthe formulation, wherein the therapeutically effective amount of the CAIcompound is 0.1 to 10 mg of the CAI compound.
 9. The method of claim 1,wherein the sterile, aqueous formulation is a time-release formulation.10. An aqueous, sterile formulation comprising a CAI compound, whereinsaid CAI compound is solubilized with excipient materials selected fromthe group consisting of: pure triglyceride oils, diglycerides,monoglycerides, mixed glycerides, lipophilic surfactants, hydrophilicsurfactants, and water-soluble cosolvents, and wherein said formulationconsists of 0.1 mg/mL to 100 mg/mL of the CAI compound.
 11. Theformulation of claim 10, wherein the CAI compound is selected from thegroup consisting of CAI free base; CAI acid addition salt, and CAIprodrug.
 12. The formulation of claim 11, wherein the CAI compound is anorganic or inorganic acid addition salt form of CAI selected from thegroup consisting of: borate, hydrobromide, hydrochloride, nitrate,phosphate, dihydrogenphosphate, sulfate, hydrogensulfate, citrate,fumarate, gluconate, glutamate, lactate, maleate, mandelate, mesylate,oxalate, succinate tartrate, valerate, benzenesulfonate, benzoate,cholate, hydroxynaphthoate, laurate, napsylate, oleate, palmoate,palmitate, salicylate, stearate, tosylate, and taurocholate.
 13. Theformulation of claim 10, further comprising any one or combination ofsubstances selected from the group consisting of: pharmaceuticallyacceptable carriers, pharmaceutically acceptable auxiliary substances,diluents, surfactants, detergents, stabilizers, excipients, carriers,and delivery-enhancing agents.
 14. The formulation of claim 10, whereinsaid formulation is selected from the group consisting of: Formulation1, Formulation 2, Formulation 3, Formulation 4, Formulation 5, andFormulation
 6. 15. The formulation of claim 10, wherein the formulationconsists of 1 mg/mL to 60 mg/mL of the CAI compound.
 16. A method forpreparing a sterile, aqueous formulation comprising a therapeuticallyeffective amount of a CAI compound, wherein the therapeuticallyeffective amount is from about 0.1 mg/mL to 100 mg/mL of the CAIcompound in the formulation; said method comprising: (a) in a solvent,mixing said CAI compound with any or combination of excipients selectedfrom the group consisting of: pure triglyceride oils, diglycerides,monoglycerides, mixed glycerides, lipophilic surfactants, hydrophilicsurfactants, and water-soluble cosolvents, to form a soluble CAIcompound; (b) adding an aqueous solution to the soluble CAI compound;and (c) subjecting the aqueous solution to heating.
 17. The method ofclaim 16, wherein the CAI compound is selected from the group consistingof CAI free base; CAI acid addition salt, and CAI prodrug.
 18. Themethod of claim 16, wherein the CAI compound is mixed with oleic acid,soybean oil, and Tween-80.
 19. The method of claim 16, wherein theheating is performed at 121° C.
 20. The method of claim 16, furthercomprising the step of subjecting the aqueous solution to pressure. 21.The method of claim 20, wherein the pressure is at 15 psi.
 22. Themethod of claim 16, wherein the solvent is absolute ethanol.
 23. Themethod of claim 16, wherein the aqueous solution is water, wherein saidmethod further comprises the step of removing the solvent.
 24. Themethod of claim 16, further comprising the step of adding any one orcombination of the following to the aqueous solution: pharmaceuticallyacceptable carriers, pharmaceutically acceptable auxiliary substances,diluents, surfactants, detergents, stabilizers, carriers, anddelivery-enhancing agents.
 25. The method of claim 16, wherein thetherapeutically effective amount of the CAI compound is 1 mg/mL to 60mg/mL.